Dicycloazaalkane derivates, preparation processes and medical uses thereof

ABSTRACT

Disclosed are new dicycloazaalkane derivates represented by general formula (I), preparation processes and pharmaceutical compositions containing them, and the uses for treatment especially for dipeptidyl peptidase inhibitor (DPP-IV), in which each substitute group of general formula (I) is as defined in specification.

The present application is the national phase application of PCTApplication No. PCT/CN2008/071014, filed May 20, 2008, which claimspriority to Chinese Patent Application No. 200810004727.1, filed Jan.23, 2008, the entireties of both of which are hereby incorporated byreferences.

FIELD

This disclosure relates to derivatives of azabicyclo alkane, methods forpreparing the same, compositions containing the same, and the usesthereof, particularly their pharmaceutical use as inhibitors ofdipeptidyl peptidase IV (DPP-IV).

BACKGROUND

Diabetes refers to a disease derived from multiple causative factors andcharacterized by elevated levels of plasma glucose or hyperglycemia,along with sugar, fat and protein metabolism disorders caused by insulinsecretion and/or the action defects. Diabetes is an ancient disease dueto the human body absolute or relative lack of insulin, which results inincreased concentrations of glucose in the blood. The glucose is largelydischarged in urine. High blood levels of glucose can cause severalproblems, such as excessive thirst, frequent urination, increasedappetite, weight loss, dizziness, fatigue, and other symptoms.

Dipeptidyl peptidase-IV (DPP-IV) is a serine protease that cleavesN-terminal dipeptides from a peptide chain containing, preferably, aproline residue in the penultimate position. Although its biologicalrole in mammalian systems has not been completely established, DPP-IV isbelieved to play an important role in neuropeptide metabolism, T-cellactivation, attachment of cancer cells to the endothelium, and the entryof HIV into lymphoid cells (WO98/19998).

More recently, it was discovered that DPP-IV is responsible forinhibiting the secretion of glucagon-like peptide (GLP)-1. Moreparticularly, DPP-IV cleaves the amino-terminal His-Ala dipeptide ofGLP-1, degrading active GLP-1(7-36)NH₂ into inactive GLP-1(9-36)NH₂(Endocrinology, 1999, 140: 5356-5363). Under the physiologicalcondition, the half-life of the whole GLP-1 in blood circulation isshort, the inactive metabolite from GLP-1 degraded by DPP-IV can combinewith GLP-1 receptor to antagonize the active GLP-1, so the physiologicalresponse to GLP-1 is shortened. The endogenous even exogenous GLP-1 canbe entirely protected by the DPP-IV inhibitor from being deactivated byDPP-IV, and the GLP-1 bioactivity can be significantly increased (5- to10-fold). Since GLP-1 is a major stimulator of pancreatic insulinsecretion and can directly effect on glucose disposal, the DPP-IVinhibitor is well useful for treating non-insulin-dependent diabetesmellitus (NIDDM) (U.S. Pat. No. 6,110,949).

SUMMARY

Accordingly, the present disclosure relates to the compounds of formula(I) or pharmaceutically acceptable salts thereof.

Wherein:

R is selected from the group consisting of alkyl, cycloalkyl, haloalkyl,aryl, heteroaryl, aminocarbonyl alkyl, amide alkyl, heterocycliccarbonyl alkyl, and heterocyclic aminoalkyl, wherein the heterocycle is5- or 6-membered heterocyclic ring further substituted with one or moregroups selected from the group consisting of alkyl, aryl, heteroaryl,haloalkyl, haloalkoxyl, hydroxyl, amino, alkylamino, amido,aminocarbonyl, cyano, alkynyl, alkoxyl, aryloxyl, aminoalkyl,hydroxyalkyl, heterocyclic alkyl, carboxylic acid, carboxylic ester andhalogen;

R¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl,heterocyclic alkyl, aryl, heteroaryl, —C(O)NR³R⁴, —C(O)R³ and —C(O)OR³,wherein the alkyl, cycloalkyl, heterocyclic alkyl, aryl or heteroaryl isfurther optionally substituted with one or more groups selected from thegroup consisting of alkyl, aryl, hydroxyl, amino, alkoxyl, aryloxyl andheterocyclic alkyl;

R² is selected from the group consisting of hydrogen and alkyl, whereinthe alkyl is further substituted with one or more groups selected fromthe group consisting of cycloalkyl and aryl;

R³ and R⁴ are each independently selected from the group consisting ofhydrogen, alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic alkyl,wherein the alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic alkyl isfurther optionally substituted with one or more groups selected from thegroup consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxyl,cycloalkoxyl, aryloxyl, heteroaryloxyl, halogen, hydroxyl, amino, cyano,hydroxyalkyl, heterocyclic alkyl, heterocyclic alkoxyl, trifluoromethyl,carboxylic acid and carboxylic ester;

R³ and R⁴ are optionally attached together with the N atom to form a 3-to 8-membered heterocyclic ring, wherein the 5- to 8-memberedheterocyclic ring further optionally contains one or more heteroatomsselected from N, O, and S, and the 3- to 8-membered ring is furtheroptionally substituted with one or more groups selected from the groupconsisting of alkyl, aryl, heteroaryl, haloalkyl, haloalkoxyl, hydroxyl,amino, cyano, alkoxyl, aryloxyl, hydroxyalkyl, heterocyclic alkyl,carboxylic acid, carboxylic ester, halogen and —NR³R⁴; and

n is an integer from 0 to 4.

Further, the present disclosure includes the compounds of formula (IA)or pharmaceutically acceptable salts thereof.

Wherein:

R is selected from the group consisting of alkyl, cycloalkyl, haloalkyl,aryl, heteroaryl, aminocarbonyl alkyl, amide alkyl, heterocycliccarbonyl alkyl, and heterocyclic aminoalkyl, wherein the heterocycle is5- or 6-membered heterocyclic ring further substituted with one or moregroups selected from the group consisting of alkyl, aryl, heteroaryl,haloalkyl, haloalkoxyl, hydroxyl, amino, alkylamino, amido,aminocarbonyl, cyano, alkynyl, alkoxyl, aryloxyl, aminoalkyl,hydroxyalkyl, heterocyclic alkyl, carboxylic acid, carboxylic ester andhalogen;

R¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl,heterocyclic alkyl, aryl, heteroaryl, —C(O)NR³R⁴, —C(O)R³ and —C(O)OR³,wherein the alkyl, cycloalkyl, heterocyclic alkyl, aryl or heteroaryl isoptionally further substituted with one or more groups selected from thegroup consisting of alkyl, aryl, hydroxyl, amino, alkoxyl, aryloxyl andheterocyclic alkyl;

R² is selected from the group consisting of hydrogen and alkyl, whereinthe alkyl is further substituted with one or more groups selected fromthe group consisting of cycloalkyl and aryl;

R³ and R⁴ are each independently selected from the group consisting ofhydrogen, alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic alkyl,wherein the alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic alkyl isfurther optionally substituted with one or more groups selected from thegroup consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxyl,cycloalkoxyl, aryloxyl, heteroaryloxyl, halogen, hydroxyl, amino, cyano,hydroxyalkyl, heterocyclic alkyl, heterocyclic alkoxyl, trifluoromethyl,carboxylic acid and carboxylic ester; and

R³ and R⁴ are optionally attached together with the N atom to form a 3-to 8-membered heterocyclic ring, wherein the 5- to 8-memberedheterocyclic ring further optionally contains one or more heteroatomsselected from N, O and S, and the 3- to 8-membered ring is furtheroptionally substituted with one or more groups selected from the groupconsisting of alkyl, aryl, heteroaryl, haloalkyl, haloalkoxyl, hydroxyl,amino, cyano, alkoxyl, aryloxyl, hydroxyalkyl, heterocyclic alkyl,carboxylic acid, carboxylic ester, halogen and —NR³R⁴.

Preferably, in the compounds of formula (I) or pharmaceuticallyacceptable salts thereof, R is the following formula.

Wherein:

R⁵ is selected from the group consisting of hydrogen, alkyl, cycloalkyl,aryl, heteroaryl and heterocyclic alkyl, wherein the alkyl, cycloalkyl,aryl, heteroaryl or heterocyclic alkyl is further optionally substitutedwith one or more groups selected from the group consisting of alkyl,cycloalkyl, aryl, heteroaryl, alkoxyl, cycloalkoxyl, aryloxyl,heteroaryloxyl, halogen, hydroxyl, amino alkylamino, cyano,hydroxyalkyl, heterocyclic alkyl, heterocyclic alkoxyl, carboxylic acidand carboxylic ester; and

R⁶ and R⁷ are each independently selected from the group consisting ofalkyl, aryl, heteroaryl, haloalkyl, haloalkoxyl, hydroxyl, amino, cyano,alkynyl, alkoxyl, aryloxyl, hydroxyalkyl, heterocyclic alkyl, carboxylicacid, carboxylic ester and halogen; and W is C, S or O, wherein C isfurther optionally substituted with R⁶ or R⁷.

Further, the present disclosure includes the compounds of formula (IB)or pharmaceutically acceptable salts thereof.

Wherein:

R is the following formula:

R¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl,heterocyclic alkyl, aryl, heteroaryl, —C(O)NR³R⁴, —C(O)R³ and —C(O)OR³,wherein the alkyl, cycloalkyl, heterocyclic alkyl, aryl or heteroaryl isfurther substituted with one or more groups selected from the groupconsisting of alkyl, aryl, hydroxyl, amino, alkoxyl, aryloxyl andheterocyclic alkyl;

R² is selected from the group consisting of hydrogen and alkyl, whereinthe alkyl is further substituted with one or more groups selected fromthe group consisting of cycloalkyl and aryl;

R³ and R⁴ are each independently selected from the group consisting ofhydrogen, alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic alkyl,wherein the alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic alkyl isfurther substituted with one or more groups selected from the groupconsisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxyl,cycloalkoxyl, aryloxyl, heteroaryloxyl, halogen, hydroxyl, amino, cyano,hydroxyalkyl, heterocyclic alkyl, heterocyclic alkoxyl, trifluoromethyl,carboxylic acid and carboxylic ester;

R³ and R⁴ are attached together with the N atom to form a 3- to8-membered heterocyclic ring, wherein the 5- to 8-membered heterocyclicring further contains one or more heteroatoms selected from N, O and S,and the 3- to 8-membered ring is further substituted with one or moregroups selected from the group consisting of alkyl, aryl, heteroaryl,haloalkyl, haloalkoxyl, hydroxyl, amino, cyano, alkoxyl, aryloxyl,hydroxyalkyl, heterocyclic alkyl, carboxylic acid, carboxylic ester,halogen, and —NR³R⁴;

R⁵ is selected from the group consisting of hydrogen, alkyl, cycloalkyl,aryl, heteroaryl and heterocyclic alkyl, wherein the alkyl, cycloalkyl,aryl, heteroaryl or heterocyclic alkyl is further substituted with oneor more groups selected from the group consisting of alkyl, cycloalkyl,aryl, heteroaryl, alkoxyl, cycloalkoxyl, aryloxyl, heteroaryloxyl,halogen, hydroxyl, amino, alkylamino, cyano, hydroxyalkyl, heterocyclicalkyl, heterocyclic alkoxyl, carboxylic acid and carboxylic ester; and

R⁶ and R⁷ are each independently selected from the group consisting ofalkyl, aryl, heteroaryl, haloalkyl, haloalkoxyl, hydroxyl, amino, cyano,alkynyl, alkoxyl, aryloxyl, hydroxyalkyl, heterocyclic alkyl, carboxylicacid, carboxylic ester and halogen; and W is C, S or O, wherein C can befurther substituted with R⁶ or R⁷.

Further, the present disclosure includes the compounds of formula (IC)or pharmaceutically acceptable salts thereof.

Wherein: R is the following formula:

R¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl,heterocyclic alkyl, aryl, heteroaryl, —C(O)NR³R⁴, —C(O)R³ and —C(O)OR³,wherein the alkyl, cycloalkyl, heterocyclic alkyl, aryl or heteroaryl isfurther substituted with one or more groups selected from the groupconsisting of alkyl, aryl, hydroxyl, amino, alkoxyl, aryloxyl andheterocyclic alkyl;

R² is selected from the group consisting of hydrogen and alkyl, whereinthe alkyl is further substituted with one or more groups selected fromthe group consisting of cycloalkyl and aryl;

R³ and R⁴ are each independently selected from the group consisting ofhydrogen, alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic alkyl,wherein the alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic alkyl isfurther substituted with one or more groups selected from the groupconsisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxyl,cycloalkoxyl, aryloxyl, heteroaryloxyl, halogen, hydroxyl, amino, cyano,hydroxyalkyl, heterocyclic alkyl, heterocyclic alkoxyl, trifluoromethyl,carboxylic acid and carboxylic ester;

R³ and R⁴ are attached together with the N atom to form a 3- to8-membered heterocyclic ring, wherein the 5- to 8-membered heterocyclicring further contains one or more heteroatoms selected from N, O and S,and the 3- to 8-membered ring is further substituted with one or moregroups selected from the group consisting of alkyl, aryl, heteroaryl,haloalkyl, haloalkoxyl, hydroxyl, amino, cyano, alkoxyl, aryloxyl,hydroxyalkyl, heterocyclic alkyl, carboxylic acid, carboxylic ester,halogen and —NR³R⁴;

R⁵ is selected from the group consisting of hydrogen, alkyl, cycloalkyl,aryl, heteroaryl and heterocyclic alkyl, wherein the alkyl, cycloalkyl,aryl, heteroaryl or heterocyclic alkyl is further substituted with oneor more groups selected from the group consisting of alkyl, cycloalkyl,aryl, heteroaryl, alkoxyl, cycloalkoxyl, aryloxyl, heteroaryloxyl,halogen, hydroxyl, amino, alkylamino, cyano, hydroxyalkyl, heterocyclicalkyl, heterocyclic alkoxyl, carboxylic acid and carboxylic ester; and

R⁶ and R⁷ are each independently selected from the group consisting ofalkyl, aryl, heteroaryl, haloalkyl, haloalkoxyl, hydroxyl, amino, cyano,alkynyl, alkoxyl, aryloxyl, hydroxyalkyl, heterocyclic alkyl, carboxylicacid, carboxylic ester and halogen; and W is C, S or O, wherein C can befurther substituted with R⁶ or R⁷.

This disclosure provides compounds of formula (I) or pharmaceuticallyacceptable salts, wherein the salts are obtained by reaction of thecompounds of formula (I) with the acids selected from the groupconsisting of hydrochloric acid, p-toluenesulfonic acid, tartaric acid,maleic acid, lactic acid, methanesulfonic acid, sulfuric acid,phosphoric acid, citric acid, acetic acid and trifluoroacetic acid.Preferred acids are p-toluenesulfonic acid, hydrochloric acid, tartaricacid, and trifluoroacetic acid.

In a particularly preferred embodiment, the compounds of formula (I) orpharmaceutically acceptable salts include:

Example No. Structure Name 1

cis-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2- oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2- carboxylic acid dimethylamide hydrochloride 2

cis-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxoethylamino]-hexahydro-cyclopenta[c] pyrrole-2-carboxylic acid methylester hydrochloride 3

cis-(S)-1-{2-[2-(2-Hydroxy-acetyl)- octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2- carbonitrile hydrochloride 4

cis-(S)-1-{2-[2-(Piperidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5- ylamino]-acetyl}-pyrrolidine-2-carbonitrile hydrochloride 5

cis-(S)-1-[2-(2-Acetyl-octahydro-cyclopenta[c]pyrrol-5-ylamino)-acetyl]- pyrrolidine-2-carbonitrilehydrochloride 6

cis-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)- 2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acid isopropylamide hydrochloride 7

cis-(S)-1-{2-[2-(Morpholine-4-carbonyl)-octahydro-cyclopenta[c]pyrrol-5- ylamino]-acetyl}-pyrrolidine-2-carbonitrile hydrochloride 8

cis-(S)-1-{2-[2-(Pyrrolidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5- ylamino]-acetyl}-pyrrolidine-2-carbonitrile hydrochloride 9

cis-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2- oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2- carboxylic acid dimethylamide trifluoroacetate10

trans-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)- 2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acid dimethylamide trifluoroacetate 11

5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro- cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide p-toluenesulfonate 12

5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro- cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide p-toluenesulfonate 13

5-[2-((2S,4S)-2-Cyano-4-fluoro- pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl- hexahydrocyclopenta[c]pyrrole-2- carboxamide tartrate14

5-Benzyl-5-[2-((2S,4S)-2-cyano-4- fluoropyrrolidin-1-yl)-2-oxo-ethylamino]- hexahydrocyclopenta[c]pyrrole-2- carboxylic aciddimethylamide tartrate 15

5-Cyclohexylmethyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]- hexahydrocyclopenta[c]pyrrole-2-carboxylic acid dimethylamide tartrate 16

5-Cyclopentyl-5-[2-((S)-2-cyano- pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydrocyclopenta[c]pyrrole-2- carboxylic acid dimethylamide tartrate17

5-Benzyl-5-[2-((S)-2-cyano-pyrrolidin-1- yl)-2-oxo-ethylamino]-hexahydrocyclopenta[c]pyrrole-2- carboxylic acid dimethylamide tartrate18

5-[2-((2S,4S)-2-Cyano-4-fluoro- pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl- hexahydrocyclopenta[c]pyrrole-2- carboxylicacid dimethylamide p-toluenesulfonate 19

5-Ethyl-5-[2-((S)-2-cyano-pyrrolidin-1- yl)-2-oxo-ethylamino]-hexahydrocyclopenta[c]pyrrole-2- carboxylic acid dimethylamide tartrate

Further, this disclosure relates to the compounds of formula (ID) asintermediates in the synthesis of compounds of formula (I).

Wherein:

R¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl,heterocyclic alkyl, aryl, heteroaryl, —C(O)NR³R⁴, —C(O)R³ and —C(O)OR³,wherein the alkyl, cycloalkyl, heterocyclic alkyl, aryl or heteroaryl isfurther substituted with one or more groups selected from the groupconsisting of alkyl, aryl, hydroxyl, amino, alkoxyl, aryloxyl andheterocyclic alkyl;

R³ and R⁴ are each independently selected from the group consisting ofhydrogen, alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic alkyl,wherein the alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic alkyl isfurther substituted with one or more groups selected from the groupconsisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxyl,cycloalkoxyl, aryloxyl, heteroaryloxyl, halogen, hydroxyl, amino, cyano,hydroxyalkyl, heterocyclic alkyl, heterocyclic alkoxyl, trifluoromethyl,carboxylic acid and carboxylic ester; and

R³ and R⁴ are attached together with the N atom to form a 3- to8-membered heterocyclic ring, wherein the 5- to 8-membered heterocyclicring further contains one or more heteroatoms selected from N, O and S,and the 3- to 8-membered ring is further substituted with one or moregroups selected from the group consisting of alkyl, aryl, heteroaryl,haloalkyl, haloalkoxyl, hydroxyl, amino, cyano, alkoxyl, aryloxyl,hydroxyalkyl, heterocyclic alkyl, carboxylic acid, carboxylic ester,halogen, and —NR³R⁴.

Furthermore, this disclosure relates to a process for the preparation ofcompounds of formula (ID), comprising the following steps of:

reacting starting materialtert-5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acid ester withtrifluoroacetic acid in a suitable solvent such as dichloromethane, uponcooling by an ice-water bath, to obtainhexahydro-cyclopenta[c]pyrrol-5-one trifluoroacetate; and

reacting hexahydro-cyclopenta[c]pyrrol-5-one trifluoroacetate with thecorresponding acyl chloride or ester in the presence of a base to obtainthe compound of formula (ID).

This disclosure relates to the process for the preparation of compoundsof formula (IB), comprising the following step:

reacting the intermediate of formula (ID), in a suitable solvent such asmethanol or ethanol, with an amine, substituted sodium borohydride, anda suitable base such as triethylamine to obtain the compound of formula(IB).

This disclosure relates to the process for the preparation of compoundsof formula (IC), comprising the following steps of:

heating potassium tert-butoxide and a solution ofmethyltriphenylphosphonium iodide in toluene; then adding theintermediate of formula (ID) at room temperature to obtain an azabicycloalkenyl compound;

reacting the azabicyclo alkenyl compound, in a suitable solvent such asdichloromethane, with trimethylsilyl cyanide in the presence of silverperchlorate at room temperature to obtain an azabicyclo cyano compound;

reacting the azabicyclo cyano compound in a suitable solvent such asethanol with a suitable acid at room temperature to obtain an azabicycloamine compound; and

reacting the azabicyclo amine compound with a halo substituted compoundin the presence of an alkaline solvent such as N,N-dimethylformamide toobtain compound of formula (IC).

This disclosure relates to the process for the preparation of compoundsof formula (IB), comprising the following steps of:

reacting the intermediate of formula (ID) with tosylmethyl isocyanide ina suitable solvent such as ethylene glycol dimethyl ether via anisocyanide reaction to obtain an azabicyclo cyano compound;

reacting the azabicyclo cyano compound in a suitable solvent such astetrahydrofuran with a halo compound in the presence of lithiumhexamethyldisilazide to obtain a R² substituted azabicyclo cyanocompound;

hydrolyzing the R² substituted azabicyclo cyano compound in the presenceof an acid to obtain a R² substituted azabicyclo carboxyl compound; orreacting the R² substituted azabicyclo cyano compound with a reductantsuch as DIBAL-H in a suitable solvent such as dichloromethane uponcooling by an ice-water bath, to obtain an aldehyde compound; reactingthe aldehyde compound in a solvent mixture of tetradrofuran/water withsodium dihydrogen phosphate, sodium chlorite and 2-methyl-2-butene, uponcooling by an ice-water bath, to obtain a R² substituted azabicyclocarboxyl compound;

reacting the R² substituted azabicyclo carboxyl compound with ethylchloroformate in the presence of a base such as triethylamine via azidoreaction to obtain an azabicyclo azido compound;

heating the azabicyclo azido compound in a suitable solvent such astoluene followed by stirring in an acidic solution; neutralizing thereaction solution to pH showing alkalescence to obtain a R² substitutedazabicyclo amine compound; and

reacting the R² substituted azabicyclo amine compound with a halosubstituted compound in the presence of an alkaline solvent such asN,N-dimethylformamide to obtain the compound of formula (IB).

This disclosure relates to the process for the preparation of compoundsof formula (IC), comprising the following steps of:

reacting the intermediate of formula (ID) in a suitable solvent such astetrahydrofuran with a reductant such as lithium tri-tert-butoxyaluminumhydride upon cooling by an ice-water bath to obtain an azabicyclohydroxyl compound;

reacting the azabicyclo hydroxyl compound in a suitable solvent such asdichloromethane with an alkaline agent such as triethylamine andmethylsulfonyl chloride to obtain an azabicyclo methyl sulfonic acidcompound;

heating the azabicyclo methyl sulfonic acid compound andphthalimide-potassium in the presence of an alkaline agent such asN,N-dimethylformamide to obtain a phthalimide substituted azabicyclocompound;

heating the phthalimide substituted azabicyclo compound and hydrazine ina suitable solvent such as ethanol to obtain an azabicyclo aminecompound; and

heating the azabicyclo amine compound and a halo compound in a suitablesolvent such as dichloromethane to obtain compound of formula (IC).

Preferably, in the preparation processes described above, R is thefollowing formula:

R⁵ is selected from the group consisting of hydrogen, alkyl, cycloalkyl,aryl, heteroaryl and heterocyclic alkyl, wherein the alkyl, cycloalkyl,aryl, heteroaryl or heterocyclic alkyl is further substituted with oneor more groups selected from the group consisting of alkyl, cycloalkyl,aryl, heteroaryl, alkoxyl, cycloalkoxyl, aryloxyl, heteroaryloxyl,halogen, hydroxyl, amino, alkylamino, cyano, hydroxyalkyl, heterocyclicalkyl, heterocyclic alkoxyl, carboxylic acid and carboxylic ester.

R⁶ and R⁷ are each independently selected from the group consisting ofalkyl, aryl, heteroaryl, haloalkyl, haloalkoxyl, hydroxyl, amino, cyano,alkynyl, alkoxyl, aryloxyl, hydroxyalkyl, heterocyclic alkyl, carboxylicacid, carboxylic ester and halogen; and W is C, S or O, wherein C can befurther substituted with R⁶ or R⁷.

The purified compounds of formula (IB) and formula (IC) are furtherreacted with acids in the solvent of methanol, dichloromethane or ethylacetate to obtain the acid addition salts.

Furthermore, this disclosure relates to a pharmaceutical compositioncomprising the compounds of the present disclosure or salts thereof inan effective therapeutic dose, and a pharmaceutically acceptablecarrier.

Furthermore, this disclosure relates to a use of the compounds of thepresent disclosure or pharmaceutical acceptable salts in the preparationof a medicament as a dipeptidyl peptidase (DPP-IV) inhibitor.

This disclosure also relates to the compounds of formula (I) orpharmaceutically acceptable salts thereof, wherein the compounds offormula (I) may be present in free form or in the form of acid additionsalts which are pharmaceutically acceptable non-toxic. Thepharmaceutically acceptable salts include hydrochloride,p-toluenesulfonate, tartarate, maleate, lactate, methanesulfonate,sulfate, phosphate, citrate, acetate and trifluoroacetate, preferablyp-toluenesulfonate, hydrochloride, tartarate and trifluoroacetate.

DETAILED DESCRIPTION OF THE DISCLOSURE

Unless otherwise stated, the following terms used in the specificationand claims have the meanings discussed below.

The term “alkyl” refers to a saturated aliphatic hydrocarbon groupincluding C₁-C₂₀ straight chain and branched chain groups. Preferably analkyl group is a moderate size alkyl having 1 to 10 carbon atoms, e.g.,methyl, ethyl, propyl, 2-propyl, n-butyl, iso-butyl, tert-butyl, pentyl,and the like. More preferably, it is a lower alkyl having 1 to 4 carbonatoms, e.g., methyl, ethyl, propyl, 2-propyl, n-butyl, iso-butyl, ortert-butyl, and the like. The alkyl group may be substituted orunsubstituted. When substituted, the substituent group(s) is preferablyhalo, hydroxyl, lower alkoxy, aryl, aryloxy, heteroaryl, heterocyclicalkyl, —C(O)R³ and —C(O)NR³R⁴.

The term “cycloalkyl” refers to a 3- to 8-membered all-carbon monocyclicring, an all-carbon 5-membered/6-membered or 6-membered/6-membered fusedbicyclic ring or a multicyclic fused ring (a “fused” ring system meansthat each ring in the system shares an adjacent pair of carbon atomswith other ring in the system) group wherein one or more rings maycontain one or more double bonds, but none of the rings has a completelyconjugated pi-electron system. Examples of cycloalkyl groups arecyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, chcyclohexyl,cyclohexadienyl, adamantyl, cycloheptyl, cycloheptatrienyl, and thelike. The cycloalkyl group may be substituted or unsubstituted. Whensubstituted, the substituent group(s) is preferably one or moreindependently selected from the group consisting of lower alkyl,trihaloalkyl, halo, hydroxy, lower alkoxy, aryl (optionally substitutedwith one or more groups which each independently is halo, hydroxy, loweralkyl or lower alkoxy groups), aryloxy (optionally substituted with oneor more groups which each independently is halo, hydroxy, lower alkyl orlower alkoxy groups), 6-membered heteroaryl (having 1 to 3 nitrogenatoms on the ring, the carbons on the ring being optionally substitutedwith one or more groups which each independently is halo, hydroxy, loweralkyl or lower alkoxy groups), 5-membered heteroaryl (having 1 to 3heteroatoms selected from the group consisting of nitrogen, oxygen andsulfur, the carbon and nitrogen atoms of the group being optionallysubstituted with one or more groups which each independently is halo,hydroxy, lower alkyl or lower alkoxy groups), 5- or 6-memberedheterocyclic alkyl (having 1 to 3 heteroatoms selected from the groupconsisting of nitrogen, oxygen and sulfur, the carbon and nitrogen (ifpresent) atoms of the group being optionally substituted with one ormore groups which each independently is halo, hydroxy, lower alkyl orlower alkoxy groups), mercapto, (lower alkyl) thio, arylthio (optionallysubstituted with one or more groups which each independently is halo,hydroxy, lower alkyl or lower alkoxy groups), cyano, acyl, thioacyl,O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido,N-amido, nitro, N-sulfonamido, S-sulfonamido, C(O)R³, C(O)NR³R⁴ and—C(O)OR³

The term “alkenyl” refers to an alkyl group as defined above having atleast 2 carbon atoms and at least one carbon-carbon double bond.Representative examples include, but are not limited to ethenyl,1-propenyl, 2-propenyl, 1-, 2-, 3-butenyl, and the like.

The term “alkynyl” refers to an alkyl group as defined above having atleast 2 carbon atoms and at least one carbon-carbon triple bond.Representative examples include, but are not limited to ethynyl,1-propynyl, 2-propynyl, 1-, 2-, 3-butynyl, and the like.

The term “aryl” refers to groups having at least one aromatic ring,i.e., having a conjugated pi-electron system, including all-carboncyclic aryl, heteroaryl and biaryl group. The aryl group may beoptionally substituted with one or more groups each independentlyselected from the group consisting of halo, trihalomethyl, hydroxy, SR,nitro, cyano, alkoxyl and alkyl.

The term “heteroaryl” refers to an aryl having 1 to 3 heteroatomsselected from the group consisting of N, O, and S as ring atoms, theremaining ring atoms being C. The ring is 5- or 6-membered ring. Theexamples of heteroaryl groups include furyl, thienyl, pyridyl, pyrrolyl,N-alkyl pyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, and the like.

The term “heterocyclic alkyl” refers to a monocyclic or fused ring groupof 5 to 9 ring atoms, wherein one, or two ring heteroatoms are selectedfrom the group consisting of N, O, and S(O)_(n) (n is integer from 0 to2), the remaining ring atoms are C, in addition, the ring may also haveone or more double bonds, but not have a completely conjugatedpi-electron system. The unsubstituted heterocyclic alkyl includes, butis not limited to pyrrolidyl, piperidyl, piperazinyl, morpholinyl,thiomorpholinyl, homopiperazinyl, and the like. The heterocyclic alkylmay be substituted or unsubstituted. When substituted, the substituentgroup is preferably one or more, more preferably one, two, or three,further more preferably one or two groups, each independently selectedfrom the group consisting of lower alkyl, trihaloalkyl, halo, hydroxy,lower alkoxy, cyano and acyl. Preferably, the heterocyclic alkyl isoptionally substituted with one or two groups independently selectedfrom the group consisting of halo, lower alkyl, trihaloalkyl, hydroxy,mercapto, cyano, N-amido, and carboxy.

The term “hydroxy” refers to an —OH group.

The term “alkoxyl” refers to both an —O-(alkyl) and an —O-(unsubstitutedcycloalkyl) group. Representative examples include, but are not limitedto, methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy,cyclopentyloxy, cyclohexyloxy, and the like.

The term “haloalkoxy” refers to an —O-(haloalkyl). Representativeexamples include, but are not limited to, trifluoromethoxy,tribromomethoxy, and the like.

The term “aryloxyl” refers to both an —O-aryl and an —O-heteroarylgroup, wherein the aryl and heteroaryl are as defined above.Representative examples include, but are not limited to, phenoxy,pyridinyloxy, furanyloxy, thienyloxy, pyrimidinyloxy, pyrazinyloxy, andthe like, and derivatives thereof.

The term “mercapto” refers to a —SH group.

The term “alkylthio” refers to a —S-(alkyl) and a —S-(unsubstitutedcycloalkyl) group. Representative examples include, but are not limitedto, methylthio, ethylthio, propylthio, butylthio, cyclopropylthio,cyclobutylthio, cyclopentylthio, cyclohexylthio, and the like.

The term “arylthio” refers to a —S-aryl and a —S-heteroaryl group,wherein the aryl and heteroaryl are as defined above. Representativeexamples include, but are not limited to, e.g., phenylthio,pyridinylthio, furanylthio, thienylthio, pyrimidinylthio, and the like,and derivatives thereof.

The term “acyl” refers to a —C(O)—R″ group, where R″ is selected fromthe group consisting of hydrogen, lower alkyl, trihalomethyl,unsubstituted cycloalkyl, aryl (optionally substituted with one or more,preferably one, two, or three substituents selected from the groupconsisting of lower alkyl, trihalomethyl, lower alkoxy and halo groups),heteroaryl (bonded through a ring carbon) (optionally substituted withone or more, preferably one, two, or three substitutents selected fromthe group consisting of lower alkyl, trihaloalkyl, lower alkoxy and halogroups), and heteroalicyclic (bonded through a ring carbon) (optionallysubstituted with one or more, preferably one, two, or three substituentsselected from the group consisting of lower alkyl, trihaloalkyl, loweralkoxy and halo groups). Representative acyl groups include, but are notlimited to, acetyl, trifluoroacetyl, benzoyl, and the like.

The term “thioacyl” refers to a —C(S)—R″ group, wherein R″ is as definedabove.

The term “acetyl” refers to a —C(O)CH₃ group.

The term “halo” refers to fluoro, chloro, bromo, or iodo, preferablyfluoro or chloro.

The term “trifluoromethyl” refers to a —CF3 group.

The term “cyano” refers to a —C≡N group.

The term “amino” refers to a —NH₂ group.

The term “carboxylic acid” refers to a —COOH group.

The term “carboxylic ester” refers to a —COOR group, wherein R is alkylor cycloalkyl.

The term “hydroxyl alkyl” refers to a —(CH₂)_(r)OH group, wherein r isan integer from 1 to 4.

The term “optional” or “optionally” means that the subsequentlydescribed event or circumstance may or may not occur, and that thedescription includes instances where the event or circumstance may ormay not occur. For example, “heterocycle group optionally substitutedwith an alkyl group” means that the alkyl may or may not be present, andthe description includes situations where the heterocyclic group issubstituted with an alkyl group and situations where the heterocyclicgroup is not substituted with the alkyl group.

The term “pharmaceutical composition” refers to a mixture of one or moreof the compounds described herein, or physiologically/pharmaceuticallyacceptable salts or prodrugs thereof, with other chemical components,such as physiologically/pharmaceutically acceptable carriers andexcipients. The purpose of a pharmaceutical composition is to facilitateadministration of a compound to an organism.

Synthesis Method of the Disclosure Compound

Scheme I includes the steps of: reactingtert-butyl-5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acid esterwith trifluoroacetic acid in a suitable solvent such as dichloromethane,upon cooling by an ice-water bath, to obtainhexahydro-cyclopenta[c]pyrrol-5-one trifluoroacetate; reactinghexahydro-cyclopenta[c]pyrrol-5-one trifluoroacetate with an acylchloride or ester in the presence of a base to obtain the compound offormula (ID); and reacting the intermediate of formula (ID), in asuitable solvent such as methanol or ethanol, with corresponding amines,substituted sodium borohydride and a suitable base such as triethylamineto obtain the compound of formula (IB) at room temperature.

Scheme II includes the steps of: heating potassium tert-butoxide and asolution of methyltriphenylphosphonium iodide in toluene followed byaddition of the intermediate of formula (ID) at room temperature toobtain an azabicyclo alkenyl compound; reacting the azabicyclo alkenylcompound, in a suitable solvent such as dichloromethane, withtrimethylsilyl cyanide in the presence of silver perchlorate at roomtemperature to obtain an azabicyclo cyano compound; reacting theazabicyclo cyano compound in a suitable solvent such as ethanol with asuitable acid such as hydrochloric acid at room temperature to obtain anazabicyclo amine compound; and reacting the azabicyclo amine compoundwith a halo substituted compound in the presence of an alkaline solventsuch as N,N-dimethylformamide to obtain compound of formula (IC).

Scheme III includes the steps of: reacting the intermediate of formula(ID) with tosylmethyl isocyanide in a suitable solvent such as ethyleneglycol dimethyl ether via an isocyanide reaction to obtain an azabicyclocyano compound; reacting the azabicyclo cyano compound in a suitablesolvent such as tetrahydrofuran with a halo compound in the presence oflithium hexamethyldisilazide to obtain a R² substituted azabicyclo cyanocompound; hydrolyzing the R² substituted azabicyclo cyano compound inthe presence of an acid to obtain a R² substituted azabicyclo carboxylcompound or reacting the R² substituted azabicyclo cyano compound with areductant such as DIBAL-H in a suitable solvent such as dichloromethaneupon cooling by an ice-water bath, to obtain an aldehyde compound;reacting the aldehyde compound in a solvent mixture oftetradrofuran/water with sodium dihydrogen phosphate, sodium chloriteand 2-methyl-2-butene, upon cooling by an ice-water bath, to obtain a R²substituted azabicyclo carboxyl compound; reacting the R² substitutedazabicyclo carboxyl compound with ethyl chloroformate in the presence ofa base such as triethylamine via azido reaction to obtain an azabicycloazido compound; heating the azabicyclo azido compound in a suitablesolvent such as toluene followed by stirring in an acidic solution;neutralizing the reaction solution to pH showing alkalescence to obtaina R² substituted azabicyclo amine compound; and reacting the R²substituted azabicyclo amine compound with a halo substituted compoundin the presence of an alkaline solvent such as N,N-dimethylformamide toobtain the compound of formula (IB).

Scheme IV includes the steps of: reacting the intermediate of formula(ID) in a suitable solvent such as tetrahydrofuran with a reductant suchas lithium tri-tert-butoxyaluminum hydride upon cooling by an ice-waterbath to obtain an azabicyclo hydroxyl compound; reacting the azabicyclohydroxyl compound in a suitable solvent such as dichloromethane with analkaline agent such as triethylamine and methylsulfonyl chloride toobtain an azabicyclo methyl sulfonic acid compound; heating theazabicyclo methyl sulfonic acid compound and phthalimide-potassium inthe presence of an alkaline agent such as N,N-dimethylformamide toobtain a phthalimide substituted azabicyclo compound; heating thephthalimide substituted azabicyclo compound and hydrazine in a suitablesolvent such as ethanol to obtain an azabicyclo amine compound; andheating the azabicyclo amine compound and a halo compound in a suitablesolvent such as dichloromethane to obtain compound of formula (IC).

The purified compounds of formula (IB) and formula (IC) are furtherreacted with acids in the solvent of methanol, dichloromethane or ethylacetate to obtain the acid addition salts.

Furthermore, this disclosure relates to a pharmaceutical compositioncomprising the compounds of the present disclosure or salts thereof inan effective therapeutic dose, and a pharmaceutically acceptablecarrier; and a use of the compounds of the present disclosure orpharmaceutical acceptable salts in the preparation of a medicament as adipeptidyl peptidase (DPP-IV) inhibitor. In other words, this disclosurealso provides a pharmaceutical composition comprising the abovementioned compounds in an effective therapeutic dose, as well as theiruse in the preparation of a medicament as a dipeptidyl peptidase(DPP-IV) inhibitor.

Specific Implementation Methods

The following examples serve to illustrate the disclosure, but theexamples should not be considered as limiting the scope of thedisclosure.

EXAMPLES

The structures of all compounds were identified by nuclear magneticresonance (¹H NMR) and mass spectrometry (MS). ¹H NMR chemical shifts(6) were recorded in ppm (10⁻⁶). NMR was performed on a BrukerAVANCE-400 spectrometer. The suitable solvents weredeuterated-chloroform (CDCl₃), deuterated-dimethyl sulfoxide (DMSO-d₆)and deuterated-methanol (CD₃OD) with tetramethylsilane (TMS) as aninternal standard and chemical shifts were recorded in ppm (10⁻⁶).

MS was determined by a FINNIGA N LCQ Ad (ESI) mass spectrometer.

The average of inhibitory rate of kinase and IC₅₀ was determined by aNovoStar ELIASA (BMG Co. German)

Thin-layer silica gel was Yantai Huanghai HSGF254 or Qingdao GF254silica gel plate.

Column chromatography studies generally used Yantai Huanghai 200-300mesh silica gel as a carrier.

DMSO-D₆: deuterated-dimethyl sulfoxide.

CDCl₃: deuterated-chloroform.

CD₃OD: deuterated-methanol

Example 1cis-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamidehydrochloride

Step 1 Preparation of(S)-[2-(2-carbamoyl-pyrrolidin-1-yl)-2-oxo-ethyl]-carbamic acidtent-butyl ester 1b

N-tert-butyloxycarbonyl glycine 1a (5 g, 28.56 mmol) and L-prolinamide(3.25 g, 28.50 mmol) were dissolved in 75 mL of N,N-dimethylformamideand the mixture was cooled to 0° C. Then 1-hydroxybenzotriazole (11.8 g,87.3 mmol), N-ethyl-N′-(dimethylaminopropyl)-carbodiimide (11.3 g, 59mmol) and triethylamine (12.1 mL, 87.3 mmol) were added with stirring.The reaction mixture was warmed up to room temperature and stirredovernight. The reaction was monitored by thin layer chromatography (TLC)until the disappearance of the starting materials. N,N-dimethylformamidewas evaporated below 50° C. The resulting mixture was extracted withethyl acetate (200 mL×3). The combined organic extracts were washed with50 mL of saturated brine, dried over anhydrous sodium sulfate, filteredand concentrated under reduced pressure. The residue was recrystallizedfrom ethyl acetate to obtain the title compound(S)-[2-(2-carbamoyl-pyrrolidin-1-yl)-2-oxo-ethyl]-carbamic acidtert-butyl ester 1b (7.42 g, yield 95.8%) as a white powder.

MS m/z (ESI): 272.1 [M+1]

Step 2 Preparation of(S)-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethyl]-carbamic acid tert-butylester 1c

In a dry three-necked flask, 286 mL of pyridine,(S)-[2-(2-carbamoyl-pyrrolidin-1-yl)-2-oxo-ethyl]-carbamic acidtert-butyl ester 1b (13.5 g, 49.8 mmol) and imidazole (7.11 g, 104.6mmol) were added successively under nitrogen atmosphere. Then thereaction system was cooled to −35° C. followed by dropwise addition ofphosphorus oxychloride (19 mL, 204.2 mmol) with stirring. The reactionmixture was stirred for 1 hour at the same temperature. After warmed upto room temperature, the reaction mixture was stirred for another 30minutes. The mixture was evaporated to remove pyridine and the resultingmixture was diluted with ethyl acetate followed by addition of water.The mixture was extracted with ethyl acetate (200 mL×3). The combinedorganic extracts were washed with 50 mL of saturated brine, dried overanhydrous sodium sulfate, filtered and concentrated under reducedpressure. The resulting residue was purified by silica gel columnchromatography to obtain the title compound(S)-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethyl]-carbamic acid tert-butylester 1c (10.7 g, yield 84.9%) as a white powder.

MS m/z (ESI): 254.3 [M+1]

Step 3 Preparation of (S)-1-(2-amino-acetyl)-pyrrolidine-2-carbonitrilehydrochloride 1d

(S)-[2-(2-Cyano-pyrrolidin-1-yl)-2-oxo-ethyl]-carbamic acid tert-butylester 1c (13.7 g, 54.2 mmol) was dissolved in 140 mL of ether and 40 mLof water with stirring. Upon cooling by an ice-water bath, 37%hydrochloric acid (90 mL) was dropwise added and the reaction mixturewas stirred for 1 hour at the same temperature. The mixture wasconcentrated under reduced pressure and the residue diluted with etherwas filtered with a filtering centrifuge to obtain the title compound(S)-1-(2-amino-acetyl)-pyrrolidine-2-carbonitrile hydrochloride 1d (10g, yield 98%) as a white powder.

MS m/z (ESI): 154.4 [M+1]

Step 4 Preparation of hexahydro-cyclopenta[c]pyrrol-5-onetrifluoroacetate 1f

tert-Butyl 5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxylate 1e (0.32g, 1.42 mmol) was dissolved in 10 mL of dichloromethane with stirring.Upon cooling by an ice-water bath, trifluoroacetic acid (3.27 mL, 42.7mmol) was added and the reaction mixture was reacted for 30 minutes at0° C. The mixture was concentrated under reduced pressure to obtain thecrude title compound hexahydro-cyclopenta[c]pyrrol-5-onetrifluoroacetate 1f, which was directly used in the next step.

MS m/z (ESI): 126.4 [M+1]

Step 5 Preparation ofN,N-dimethyl-5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxamide 1g

Hexahydro-cyclopenta[c]pyrrol-5-one trifluoroacetate if obtained fromthe above step was dissolved in 15 mL of acetonitrile with stirring.Upon cooling by an ice-water bath, potassium carbonate (0.24 g, 1.71mmol) was added followed by dimethylcarbamic chloride (0.14 mL, 1.56mmol). The reaction mixture was warmed up to room temperature andreacted for 2 hours. The mixture was concentrated under reduced pressureand diluted with 50 mL of water. The mixture was extracted with ethylacetate (50 mL×3). The combined organic extracts were washed with 50 mLof saturated brine, dried over anhydrous magnesium sulfate, filtered andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography to obtain the title compoundN,N-dimethyl-5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxamide 1g (0.19g, yield 68.3%) as a light yellow oil.

MS m/z (ESI): 197.4 [M+1]

¹H NMR (DMSO-D₆, 400 MHz): δ 3.56 (m, 2H), 2.85 (m, 2H), 2.7 (s, 6H),2.81 (m, 2H), 2.5 (m, 2H), 2.01 (m, 2H).

Step 6 Preparation ofcis-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethylhexahydro-cyclopenta[c]pyrrole-2-carboxamide 1 h

(S)-1-(2-Amino-acetyl)-pyrrolidine-2-carbonitrile hydrochloride 1d (0.36g, 1.91 mmol) was dissolved in 20 mL of methanol with stirring, followedby addition ofN,N-dimethyl-5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxamide 1g (0.25g, 1.28 mmol) and sodium triacetoxyborohydride (1.22 g, 5.74 mmol). Thereaction mixture was reacted for 3 hours at room temperature. Themixture was concentrated and diluted with 20 mL of saturated aqueoussodium carbonate. Then the mixture was extracted with dichloromethane(20 mL×10). The combined organic extracts were washed with 10 mL ofsaturated brine, dried over anhydrous sodium sulfate, filtered andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography to obtain the title compoundcis-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethylhexahydro-cyclopenta[c]pyrrole-2-carboxamide 1 h (0.3 mg, yield 53%) asa white powder.

MS m/z (ESI): 334.5 [M+1]

Step 7cis-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamidehydrochloride

cis-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethylhexahydro-cyclopenta[c]pyrrole-2-carboxamide 1 h (200 mg, 0.687 mmol)was dissolved in 10 mL of dichloromethane. Upon cooling by an ice-waterbath, 2 mL of hydrochloric acid in ether (0.5 N) was added. The mixturewas concentrated under reduced pressure and diluted with 10 mL of ether.The resulting mixture was filtered with a filtering centrifuge to obtainthe title compoundcis-5-[2-(S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamidehydrochloride 1 (180 mg, 80%) as a white power. ¹H NMR (CD₃OD, 400 MHz):δ 4.82 (dd, 1H, J₁=4 Hz, J₂=5.2 Hz), 4.02 (dd, 2H, J₁=J₂=16.4 Hz),3.62-3.25 (m, 7H), 2.76 (s, 6H), 2.51-1.49 (m, 10H).

Example 2cis-Methyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxamidehydrochloride

Step 1 Preparation of methyl5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxamide 2a

Hexahydro-cyclopenta[c]pyrrol-5-one trifluoroacetate 1f (0.559 g, 2.34mmol) was dissolved in 20 mL of acetonitrile. Upon cooling by anice-water bath, potassium carbonate (0.646 g, 4.68 mmol) and methyl2-chloroacetate (0.22 mL, 2.8 mmol) added successively. The reactionmixture was warmed up to room temperature and stirred overnight. Themixture was concentrated under reduced pressure and diluted with 50 mLof water. The mixture was extracted with ethyl acetate (50 mL×3). Thecombined organic extracts were washed with 50 mL of saturated brine and50 mL of water successively, dried over anhydrous magnesium sulfate,filtered and concentrated under reduced pressure. The resulting residuewas purified by silica gel column chromatography to obtain the titlecompound methyl 5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxamide 2a(0.25 g, yield 58.4%) as a colorless oil.

MS m/z (ESI): 184 [M+1]

Step 2 Preparation ofcis-Methyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxamide2b

(S)-1-(2-Amino-acetyl)-pyrrolidine-2-carbonitrile hydrochloride 1d (0.43g, 2.29 mmol) was dissolved in 20 mL of methanol with stirring followedby addition of methyl 5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxamide2a (0.28 g, 1.53 mmol) and sodium triacetoxyborohydride (1.46 g, 6.88mmol). The reaction mixture was reacted for 3 hours at room temperature.The mixture was concentrated and diluted with 20 mL of saturated aqueoussodium carbonate. The mixture was extracted with dichloromethane (20mL×3). The combined organic extracts were washed with 10 mL of saturatedbrine, dried over anhydrous sodium sulfate, filtered and concentratedunder reduced pressure. The resulting residue was purified by silica gelcolumn chromatography to obtain the title compoundcis-methyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxamide2b (0.22 g, yield 41%) as a white powder.

MS m/z (ESI): 357 [M+1]

Step 3cis-Methyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxamidehydrochloride 2

cis-Methyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxamide2b was dissolved in 10 mL of ether. Upon cooling by an ice-water bath, 2mL of hydrochloric acid in ether (0.5 N) was added. The resultingmixture was filtered with a filtering centrifuge to obtain the titlecompoundcis-methyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxamidehydrochloride 2 (200 mg) as a white powder.

¹H NMR (CD₃OD, 400 MHz): δ 4.71 (m, 1H), 3.93 (m, 2H), 3.59-3.28 (m,10H), 2.64 (m, 2H), 2.34 (m, 2H), 2.17 (m, 2H), 2.08 (m, 2H).

Example 3cis-(S)-1-{2-[2-(2-Hydroxy-acetyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrilehydrochloride

Step 1 Preparation of2-(2-hydroxy-acetyl)-hexahydro-cyclopenta[c]pyrrol-5-one 3a

Hexahydro-cyclopenta[c]pyrrol-5-one trifluoroacetate 1f (764.8 mg, 3.2mmol) and 2-hydroxyl ethanoic acid (267.5 mg, 3.52 mmol) were dissolvedin 10 mL of acetonitrile. Upon cooling by an ice-water bath,hydroxyacetic acid (1.3 g, 9.6 mmol),1-ethyl-3-dimethylaminopropyl-carbodiimide hydrochloride (1.23 g, 6.4mmol) and triethylamine (1.3 mL, 9.6 mmol) were added. The ice-waterbath was removed and the reaction mixture was reacted overnight at 25°C. The mixture was concentrated and diluted with 20 mL of ethyl acetate.The mixture was filtered under reduced pressure and the filtrate waswashed with 20 mL of water. The organic phase was dried over anhydrousmagnesium sulfate, filtered under reduced pressure and concentratedunder reduced pressure. The resulting residue was purified by silica gelcolumn chromatography to obtain the title compound2-(2-hydroxy-acetyl)-hexahydro-cyclopenta[c]pyrrol-5-one 3a (0.375 g,yield 64%) as a colorless oil.

MS m/z (ESI): 184 [M+1]

Step 2 Preparation ofcis-(S)-1-{2-[2-(2-hydroxy-acetyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrile3b

2-(2-Hydroxy-acetyl)-hexahydro-cyclopenta[c]pyrrol-5-one 3a (0.375 g,2.05 mmol) and (S)-1-(2-amino-acetyl)-pyrrolidine-2-carbonitrilehydrochloride 1d (0.78 g, 4.1 mmol) were dissolved in 5 mL of methanoland 10 mL of tetrahydrofuran. After the mixture was reacted at roomtemperature for 30 minutes, sodium triacetoxyborohydride (0.87 g, 4.1mmol) was added. Then the reaction mixture was reacted overnight at roomtemperature. The mixture was concentrated under reduced pressure anddiluted with 50 mL of methanol followed by addition of potassiumcarbonate (2 g, 7 mmol). After stirred for 30 minutes, the mixture wasfiltered and the filtrate was concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography toobtain the title compoundcis-(S)-1-{2-[2-(2-hydroxy-acetyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrile3b, which was directly used in the next step.

MS m/z (ESI): 357 [M+1]

Step 3 Preparation ofcis-(S)-1-{2-[2-(2-hydroxy-acetyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrilehydrochloride 3

cis-(S)-1-{2-[2-(2-Hydroxy-acetyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrile3b was dissolved in 10 mL of ether. Upon cooling by an ice-water bath, 2mL of hydrochloric acid in ether (0.5 N) was added. The resultingmixture was filtered with a filtering centrifuge to obtain the titlecompoundcis-(S)-1-{2-[2-(2-hydroxy-acetyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrilehydrochloride 3 (100 mg) as a white powder.

Example 4

cis-(S)-1-{2-[2-(Piperidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrilehydrochloride

Step 1 Preparation of2-(piperidine-1-carbonyl)-hexahydro-cyclopenta[c]pyrrol-5-one 4a

Hexahydro-cyclopenta[c]pyrrol-5-one trifluoroacetate 1f (478 mg, 2 mmol)was dissolved in 20 mL of dichloromethane with stirring followed byaddition of ido[3-(1-piperidine-formyl)imidazole-1-methyl] (0.96 g, 3mmol) and triethylamine (0.84 mL, 6 mmol). The reaction mixture wasstirred overnight at room temperature. The reaction was quenched with 20mL of water and the mixture was extracted with dichloromethane (50mL×3). The combined organic extracts were washed with 50 mL of 10%citric acid solution and 50 mL of saturated brine successively, driedover anhydrous magnesium sulfate, filtered and concentrated underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography to obtain the title compound2-(piperidine-1-carbonyl)-hexahydro-cyclopenta[c]pyrrol-5-one 4a (0.41g, yield 87%) as a colorless oil.

MS m/z (ESI): 237 [M+1]

Step 2 Preparation ofcis-(S)-1-{2-[2-(piperidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrile4b

2-(Piperidine-1-carbonyl)-hexahydro-cyclopenta[c]pyrrol-5-one 4a (0.41g, 1.74 mmol) and (S)-1-(2-amino-acetyl)-pyrrolidine-2-carbonitrilehydrochloride 1d (0.5 g, 2.6 mmol) were dissolved in 50 mL oftetrahydrofuran followed by addition of sodium sulfate (5 g) and 0.05 mLof acetic acid. After the mixture was stirred at room temperature for 30minutes, sodium triacetoxyborohydride (1.1 g, 5.2 mmol) was added. Thereaction mixture was reacted at room temperature for 3 hours andconcentrated under reduced pressure. The mixture was diluted with 50 mLof saturated aqueous sodium carbonate and extracted with ethyl acetate(50 mL×3). The combined organic extracts were washed with 50 mL ofsaturated brine and 50 mL of water successively, dried over anhydroussodium sulfate, filtered and concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography toobtain the title compoundcis-(S)-1-{2-[2-(piperidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrile4b which was directly used in the next step.

MS m/z (ESI): 410 [M+1]

Step 3 Preparation ofcis-(S)-1-{2-[2-(piperidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrilehydrochloride 4

cis-(S)-1-{2-[2-(Piperidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrile4b was dissolved in 10 mL of ether. Upon cooling by an ice-water bath, 2mL of hydrochloric acid in ether (0.5 N) was added. The resultingmixture was filtered with a filtering centrifuge to obtain the titlecompoundcis-(S)-1-{2-[2-(piperidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrilehydrochloride 4 (0.16 g) as a white solid.

¹H NMR (CD₃OD, 400 MHz): δ 4.83 (dd, 1H, J₁=3.0 Hz, J₂=5.8 Hz), 4.09(dd, 2H, J₁=J₂=13.1 Hz), 3.70-3.30 (m, 10H), 2.72 (m, 2H), 2.47 (m, 2H),2.31-2.00 (m, 5H), 1.66-1.52 (m, 8H).

Example 5cis-(S)-1-[2-(2-Acetyl-octahydro-cyclopenta[c]pyrrol-5-ylamino)-acetyl]-pyrrolidine-2-carbonitrilehydrochloride

Step 1 Preparation of 2-acetyl-hexahydro-cyclopenta[c]pyrrol-5-one 5a

Hexahydro-cyclopenta[c]pyrrol-5-one trifluoroacetate 1f (717 mg, 3 mmol)was dissolved in 20 mL of acetonitrile followed by addition ofdi-tert-butyl dicarbonate (0.42 mL, 4.5 mmol) and triethylamine (0.98mL, 9 mmol) upon cooling by an ice-water bath. The reaction mixture wasstirred overnight at the same temperature. The mixture was concentratedand diluted with 50 mL of water. The mixture was extracted with ethylacetate (50 mL×3). The combined organic extracts were washed with 50 mLof saturated brine, dried over anhydrous magnesium sulfate, filtered andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography to obtain the title compound2-acetyl-hexahydro-cyclopenta[c]pyrrol-5-one 5a (0.36 g, yield 72%) as acolorless oil

MS m/z (ESI): 168.4 [M+1]

Step 2 Preparation ofcis-(S)-1-[2-(2-acetyl-octahydro-cyclopenta[c]pyrrol-5-ylamino)-acetyl]-pyrrolidine-2-carbonitrile5b

2-Acetyl-hexahydro-cyclopenta[c]pyrrol-5-one 5a (0.36 g, 2.15 mmol) and(S)-1-(2-amino-acetyl)-pyrrolidine-2-carbonitrile hydrochloride 1d(0.614 g, 3.23 mmol) were dissolved in 50 mL of tetrahydrofuran followedby addition of 5 g of sodium sulfate and 0.05 mL of acetic acid. Afterthe mixture was stirred at room temperature for 30 minutes, sodiumtriacetoxyborohydride (1.37 g, 6.46 mmol) was added and the mixture wasreacted for another 3 hours. The mixture was concentrated under reducedpressure and diluted with 50 mL of saturated aqueous sodium carbonate.The reaction mixture was extracted with ethyl acetate (50 mL×3). Thecombined organic extracts were washed with 50 mL of saturated brine and50 mL of water successively, dried over anhydrous magnesium sulfate,filtered and concentrated under reduced pressure. The resulting residuewas purified by silica gel column chromatography to obtain the titlecompoundcis-(S)-1-[2-(2-acetyl-octahydro-cyclopenta[c]pyrrol-5-ylamino)-acetyl]-pyrrolidine-2-carbonitrile5b which was directly used in the next step.

MS m/z (ESI): 305.5 [M+1]

Step 3 Preparation ofcis-(S)-1-[2-(2-acetyl-octahydro-cyclopenta[c]pyrrol-5-ylamino)-acetyl]-pyrrolidine-2-carbonitrilehydrochloride 5

cis-(S)-1-[2-(2-Acetyl-octahydro-cyclopenta[c]pyrrol-5-ylamino)-acetyl]-pyrrolidine-2-carbonitrile5b obtained from the above step was dissolved in 20 mL of ether. Uponcooling by an ice-water bath, 4 mL of hydrochloric acid in ether (0.5 N)was added. The resulting mixture was filtered with filtering centrifugeto obtain the title compoundcis-(S)-1-[2-(2-acetyl-octahydro-cyclopenta[c]pyrrol-5-ylamino)-acetyl]-pyrrolidine-2-carbonitrilehydrochloride 5 (0.23 g) as a white powder.

¹H NMR (CD₃OD, 400 MHz): δ 4.71 (m, 1H), 3.92 (m, 2H), 3.69-3.37 (m,7H), 2.69 (m, 2H), 2.33 (m, 2H), 2.13 (m, 2H), 2.04-2.00 (m, 5H), 1.48(m, 2H).

Example 6cis-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid isopropylamide hydrochloride

Step 1 Preparation ofN-isopropyl-5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxamide 6a

Hexahydro-cyclopenta[c]pyrrol-5-one trifluoroacetate 1f (717 mg, 3 mmol)was dissolved in 20 mL of dichloromethane with stirring upon cooling byan ice-water bath, followed by addition of isocyanate (9 mL, 9 mmol) andtriethylamine (1.7 mL, 12 mmol). The reaction mixture was reactedovernight at room temperature and diluted with 50 mL of water. Themixture was extracted with dichloromethane (50 mL×3). The combinedorganic extracts were washed with 10% citric acid solution (50 mL) and50 mL of saturated brine successively, dried over anhydrous magnesiumsulfate, filtered and concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography to obtainN-isopropyl-5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acidisopropylamide 6a (0.3 g, yield 47.6%) as a colorless oil.

MS m/z (ESI): 211 [M+1]

Step 2 Preparation ofcis-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid isopropylamide 6b

N-isopropyl-5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acidisopropylamide 6a (0.3 g, 1.43 mmol) and(S)-1-(2-amino-acetyl)-pyrrolidine-2-carbonitrile hydrochloride 1d(0.407 g, 2.14 mmol) were dissolved in 50 mL of tetrahydrofuran followedby addition of 5 g of sodium sulfate and 0.05 mL of acetic acid. Afterthe reaction mixture was stirred at room temperature for 30 minutes,sodium triacetoxyborohydride (0.9 g, 4.3 mmol) was added and the mixturewas stirred for 3 hours. The mixture was concentrated under reducedpressure and diluted with 50 mL of saturated aqueous sodium carbonate.The reaction mixture was extracted with ethyl acetate (50 mL×3). Thecombined organic extracts were washed with 50 mL of saturated brine and50 mL of water successively, dried over anhydrous magnesium sulfate,filtered and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography to obtain the titlecompoundcis-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid isopropylamide 6b, which was directly used in the next step.

MS m/z (ESI): 384 [M+1]

Step 3 Preparation ofcis-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid isopropylamide hydrochloride 6

cis-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid isopropylamide 6b obtained from the above step was dissolved in 10mL of ether. Upon cooling by an ice-water bath, 2 mL of hydrochloricacid in ether (0.5 N) was added. The resulting mixture was filtered withfiltering centrifuge to obtain the title compoundcis-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid isopropylamide hydrochloride 6 (80 mg) as a white powder.

¹H NMR (CD₃OD, 400 MHz): δ 4.70 (m, 1H), 3.92 (m, 2H), 3.76-3.32 (m,8H), 2.63-1.41 (m, 10H), 1.01 (d, 6H, J=6 Hz).

Example 7

cis-(S)-1-{2-[2-(Morpholine-4-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrilehydrochloride

Step 1 Preparation of2-(morpholine-4-carbonyl)-hexahydro-cyclopenta[c]pyrrol-5-one 7a

Hexahydro-cyclopenta[c]pyrrol-5-one trifluoroacetate 1f (574 mg, 2.4mmol) was dissolved in 20 mL of acetonitrile with stirring followed byaddition of potassium carbonate (0.397 g, 2.88 mmol), andmorpholine-4-carbonyl chloride (0.323 mL, 2.64 mmol) upon cooling by anice-water bath. The reaction mixture was reacted overnight at the sametemperature. The mixture was concentrated under reduced pressure anddiluted with 50 mL of water. The mixture was extracted with ethylacetate (50 mL×3). The combined organic extracts were washed with 50 mLof saturated brine, dried over anhydrous magnesium sulfate, filtered andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography to obtain the title compound2-(morpholine-4-carbonyl)-hexahydro-cyclopenta[c]pyrrol-5-one 7a (0.572g, yield 77.3%) as a colorless oil.

MS m/z (ESI): 239 [M+1]

Step 2 Preparation ofcis-(S)-1-{2-[2-(morpholine-4-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrile7b

2-(Morpholine-4-carbonyl)-hexahydro-cyclopenta[c]pyrrol-5-one 7a (0.64g, 2.69 mmol) and (S)-1-(2-amino-acetyl)-pyrrolidine-2-carbonitrilehydrochloride 1d (0.764 g, 4.03 mmol) were dissolved in 50 mL oftetrahydrofuran followed by addition of 5 g of sodium sulfate and 0.05mL of acetic acid. After the mixture was stirred at room temperature for30 minutes, sodium triacetoxyborohydride (1.71 g, 8.07 mmol) was addedand the mixture was reacted for another 3 hours. The mixture wasconcentrated under reduced pressure and diluted with 50 mL of saturatedaqueous sodium carbonate. The reaction mixture was extracted with ethylacetate (50 mL×3). The combined organic extracts were washed with 50 mLof saturated brine and 50 mL of water successively, dried over anhydrousmagnesium sulfate, filtered and concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography toobtain the title compoundcis-(S)-1-{2-[2-(morpholine-4-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrile7b which was directly used in the next step.

MS m/z (ESI): 376.7 [M+1]

Step 3 Preparation ofcis-1-{2-[(S)-2-(morpholine-4-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrilehydrochloride 7

cis-1-{2-[(S)-2-(Morpholine-4-carbonyl)-octahydro-cyclopenta[c]-pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrile7b obtained from the above step was dissolved in 10 mL of ether. Uponcooling by an ice-water bath, 2 mL of hydrochloric acid in ether (0.5 N)was added. The resulting mixture was filtered with a filteringcentrifuge to obtain the title compoundcis-1-{2-[(S)-2-(morpholine-4-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrilehydrochloride 7 (30 mg, yield 3%) as a white powder.

Example 8cis-(S)-1-{2-[2-(Pyrrolidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrilehydrochloride

Step 1 Preparation of2-(pyrrolidine-1-carbonyl)-hexahydro-cyclopenta[c]pyrrol-5-one 8a

Hexahydro-cyclopenta[c]pyrrol-5-one trifluoroacetate 1f (478 mg, 2 mmol)was dissolved in 20 mL of dichloromethane followed by addition ofpyrrolidine-1-carbonyl chloride (0.276 mL, 2.5 mmol) and triethylamine(0.84 mL, 6 mmol). The reaction mixture was reacted overnight at roomtemperature. The mixture was adjusted to pH 4 with a solution of 10%citric acid. The mixture was extracted with dichloromethane (50 mL×3).The combined organic extracts were washed with 50 mL of saturated brine,dried over anhydrous magnesium sulfate, filtered and concentrated underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography to obtain the title compound2-(pyrrolidine-1-carbonyl)-hexahydro-cyclopenta[c]pyrrol-5-one 8a (0.26g, yield 58.5%) as a colorless oil.

MS m/z (ESI): 223 [M+1]

Step 2 Preparation ofcis-(S)-1-{2-[2-(pyrrolidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrile8b

2-(Pyrrolidine-1-carbonyl)-hexahydro-cyclopenta[c]pyrrol-5-one 8a (0.26g, 1.17 mmol) and (S)-1-(2-amino-acetyl)-pyrrolidine-2-carbonitrilehydrochloride 1d (0.33 g, 1.75 mmol) were dissolved in 50 mL oftetrahydrofuran followed by addition of 5 g of sodium sulfate and 0.05mL of acetic acid. After the mixture was reacted at room temperature for30 minutes, sodium triacetoxyborohydride (0.75 g, 3.5 mmol) was addedand the reaction mixture was reacted for 3 hours. The mixture wasconcentrated under reduced pressure and diluted with 50 mL of saturatedaqueous sodium carbonate. The reaction mixture was extracted with ethylacetate (50 mL×3). The combined organic extracts were washed with 50 mLof saturated brine and 50 mL of water, dried over anhydrous magnesiumsulfate, filtered and concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography to obtain thetitle compoundcis-(S)-1-{2-[2-(pyrrolidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrile8b, which was directly used in the next step.

MS m/z (ESI): 396 [M+1]

Step 3 Preparation ofcis-(S)-1-{2-[2-(pyrrolidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrilehydrochloride

cis-(S)-1-{2-[2-(Pyrrolidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrile8b obtained from the above step was dissolved in 10 mL of ether. Uponcooling by an ice-water bath, 2 mL of hydrochloric acid in ether (0.5 N)was added. The resulting mixture was filtered with a filteringcentrifuge to obtain the title compoundcis-(S)-1-{2-[2-(pyrrolidine-1-carbonyl)-octahydro-cyclopenta[c]pyrrol-5-ylamino]-acetyl}-pyrrolidine-2-carbonitrilehydrochloride 8 (90 mg) as a white powder.

¹H NMR (CD₃OD, 400 MHz): δ 4.72 (m, 1H), 4.09 (m, 2H), 3.43-3.30 (m,11H), 2.62 (m, 2H), 2.35 (m, 2H), 2.18 (m, 2H), 2.08 (m, 2H), 1.77 (m,4H).

Example 9cis-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamidetrifluoroacetate

5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 1 h obtained from Example 1 was dissolved in 10 mL ofdichloromethane followed by addition of 2 mL of trifluoroacetic acidupon cooling by an ice-water bath. The reaction mixture was stirred for30 minutes. The resulting mixture was filtered with a filteringcentrifuge to obtain the title compoundcis-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamidetrifluoroacetate 9 (201 mg) as a white powder.

¹H NMR (CDCl₃, 400 MHz): δ 4.74 (t, 1H, J=5.2 Hz), 3.98 (d, 1H, J=15.6Hz), 3.79 (d, 1H, J=15.6 Hz), 3.57-3.25 (m, 7H), 2.75 (s, 6H), 2.55 (m,2H), 2.33 (m, 2H), 2.20-2.08 (m, 4H), 1.74 (m, 2H).

Example 10trans-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamidetrifluoroacetate

Step 1 Preparation ofcis-5-hydroxy-hexahydro-cyclopenta[c]pyrrole-2-carboxamideN,N-dimethylamide 10a

In a dry three-necked flask,N,N-dimethyl-5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxamide 1g (1.58g, 8.06 mmol) was dissolved in 30 mL of tetrahydrofuran with stirringunder nitrogen atmosphere. The mixture was cooled to −25° C. followed bydropwise addition of a solution of lithium tri-tert-butoxyaluminiumhydride (2.45 g, 9.6 mmol) in 30 mL of tetrahydrofuran. After thereaction mixture was reacted for 2.5 hours at the same temperature, thereaction was quenched with water. The mixture was diluted with 20 mL ofsaturated aqueous ammonium chloride and warmed up to room temperature.The layers were separated and then the aqueous layer was extracted withdichloromethane (50 mL×3). The combined organic extracts were washedwith 50 mL of saturated brine, dried over anhydrous magnesium sulfate,filtered and concentrated under reduced pressure. The resulting residuewas purified by silica gel column chromatography to obtain the titlecompound cis-5-hydroxy-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acidN,N-dimethylamide 10a (1.27 g, yield 80%) as a colorless oil.

MS m/z (ESI): 199 [M+1]

Step 2 Preparation of cis-methanesulfonic acid2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrol-5-yl ester 10b

In a dry one-necked flask,cis-5-hydroxy-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 10a (1.69 g, 8.5 mmol) was dissolved in 30 mL ofdichloromethane with stirring under nitrogen atmosphere. Upon cooling byan ice-salt bath to −5° C.˜0° C., triethylamine (1.66 mL, 14.45 mmol)and methanesulfonyl chloride (2.2 g, 21.74 mmol) were addedsuccessively. The reaction mixture was stirred for 30 minutes and warmedup to room temperature. After the reaction mixture was reacted for 2hours, the mixture was concentrated under reduced pressure and dilutedwith 20 mL of water. The reaction mixture was extracted with ethylacetate (50 mL×6). The combined organic extracts were washed with 50 mLof saturated brine, dried over anhydrous magnesium sulfate, filtered andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography to obtain the title compoundcis-methanesulfonic acid2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrol-5-yl ester 10b (1.94g, yield 83%) as a white solid.

MS m/z (ESI): 277 [M+1]

Step 3 Preparation oftrans-5-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 10c

In a dry one-necked flask, methanesulfonic acid2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrol-5-yl ester 10b (1 g,3.6 mmol) was added in 20 mL of N,N-dimethylformamide and phthalimidepotassium salt (993 mg, 5.4 mmol) with stirring under nitrogenatmosphere. The reaction mixture was warmed up to 70° C. and reacted for3 hours. The mixture was concentrated under reduced pressure to removeN,N-dimethylformamide and the residue was diluted with 20 mL of water.The mixture was extracted with ethyl acetate (50 mL×3). The combinedorganic extracts were washed with 50 mL of saturated brine, dried overanhydrous magnesium sulfate, filtered and concentrated under reducedpressure to obtain the title compoundtrans-5-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 10c (1.06 g, yield 90%) as a white solid which wasdirectly used in the next step.

MS m/z (ESI): 328 [M+1]

Step 4 Preparation oftrans-5-amino-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acidN,N-dimethylamide 10d

In a one-necked flask,trans-5-(1,3-Dioxo-1,3-dihydro-isoindol-2-yl)-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 10c was dissolved in 20 mL of ethanol (95%) withstirring followed by addition of hydrazine (490 mg, 15.3 mmol). Thereaction mixture was heated to reflux for 8 hours, and then cooled toroom temperature. The mixture was filtered and the filtrate wasconcentrated under reduced pressure to obtain a white solid. Theresulting solid was dissolved in 25 mL of methanol, filtered and thenthe filtrate was concentrated under reduced pressure. The resultingresidue was purified by alumina base column chromatography to obtain thetitle compound trans-5-amino-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid N,N-dimethylamide 10d (290 mg, yield 48%) as a colorless oil.

MS m/z (ESI): 198 [M+1]

Step 5 Preparation oftrans-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamide10e

In a dry one-necked flask, trans-1-(2-chloro-ethyl)-pyrrole-2-cyano (334mg, 1.94 mmol) and a solution oftrans-5-amino-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acidN,N-dimethylamide 10d (290 mg, 1.46 mmol) was dissolved in 20 mL ofdichloromethane. The reaction mixture was heated to reflux for 48 hours.The mixture was concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography to obtain thetitle compoundtrans-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamide10e, which was directly used in the next step.

Step 6 Preparation oftrans-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamidetrifluoroacetate 10

trans-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamide10e obtained from the above step was dissolved in 10 mL ofdichloromethane with stirring followed by addition of 2 mL oftrifluoroacetic acid. The reaction mixture was stirred for 30 minutes toobtain the title compoundtrans-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamidetrifluoroacetate 10 (201 mg) as a white solid.

MS m/z (ESI): 334 [M+1]

¹H NMR (CDCl₃, 400 MHz): δ 4.65 (m, 1H), 3.93 (d, 1H, J=15.2 Hz), 3.74(d, 1H, J=15.2 Hz), 3.69-3.19 (m, 7H), 2.77 (s, 6H), 2.18-1.96 (m, 10H).

Example 115-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide p-toluenesulfonate

Step 1 Preparation of5-methylene-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

Potassium tert-butoxide (7.17 g, 0.064 mol) andmethyltriphenylphosphonium iodide (25.8 g, 0.064 mol) were dissolved in150 mL of toluene under nitrogen atmosphere. The reaction mixture washeated to reflux for 3 hours, and cooled to room temperature. A solutionof N,N-dimethyl-5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxylate 1g(5.0 g, 0.0255 mol) in toluene was added. The reaction mixture wasstirred for 30 minutes. The reaction was monitored by TLC until thedisappearance of the starting materials. The mixture was diluted with 30mL of water and 30 mL of saturated brine and extracted with ethylacetate (200 mL×4). The combined organic extracts were dried overanhydrous magnesium sulfate, filtered and concentrated under reducedpressure. The resulting residue was purified by silica gel columnchromatography to obtain the title compound5-methylene-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 11a (4.0 g, yield 80%) as a light yellow oil.

MS m/z (ESI): 195.2 [M+1]

Step 2 Preparation of5-isocyano-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

5-Methylene-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 11a (1.6 g, 8.23 mmol) was dissolved in 30 mL ofdichloromethane with stirring followed by addition of trimethylsilylcyanide (4.08 g, 41.2 mmol) and silver perchlorate (5.12 g, 24.7 mmol)under nitrogen atmosphere. The reaction mixture was stirred overnight atroom temperature and then some of the reaction mixture was treated withsaturated aqueous sodium carbonate. The reaction was monitored by TLC,which showed that there remained most of the starting materials. Thenthe mixture was treated with 20 mL of saturated aqueous sodium carbonateupon cooling by an ice-water bath, and the reaction released heat. After10 minutes, the mixture was filtered to remove the residue. Theseparated aqueous phase was extracted with ethyl acetate (50 mL×4). Thecombined organic extracts were dried over anhydrous magnesium sulfate,filtered and concentrated under reduced pressure. The resulting residuewas purified by silica gel column chromatography to obtain5-isocyano-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 11b (0.33 g, yield 18.1%) as a light yellow oil.

GC-MS: 221.1 [M⁺]

¹H NMR (CDCl₃, 400 MHz): δ 3.31 (m, 4H), 2.97 (m, 2H), 2.84 (s, 6H),2.32 (m, 2H), 1.52 (s, 3H), 1.46 (m, 2H).

Step 3 Preparation of5-amino-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

To a solution of5-isocyano-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 11b (0.388 g, 1.75 mmol) in 15 mL of ethanol was added0.38 mL of hydrochloric acid (6 N) upon cooling by an ice-water bath.The reaction mixture was warmed up to room temperature and stirred for1.5 hours. The reaction was monitored by TLC until the disappearance ofthe starting materials. The mixture was quenched with 20 mL of saturatedaqueous sodium carbonate. The mixture was extracted with dichloromethane(50 mL×3). The combined organic extracts were evaporated to dryness. Theresulting residue was purified by silica gel column chromatography toobtain the title compound5-amino-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 11c (0.28 g, yield 75.7%) as a yellow oil.

MS m/z (ESI): 212.2 [M+1]

Step 4 Preparation of5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

5-Amino-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 11c (200 mg, 0.92 mmol) was dissolved in 8 mL ofN,N-dimethylformamide, followed by addition of1-(2-chloro-acetyl)-pyrrolidine-2-cyano (175 mg, 1.02 mmol). Thereaction mixture was reacted overnight at room temperature. The mixturewas concentrated under reduced pressure at 40˜50° C. to removeN,N-dimethylformamide. The resulting residue was purified by silica gelcolumn chromatography to obtain the title compound5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 11d (0.135 g, yield 42.3%) as a colorless oil. And5-amino-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 11c (130 mg) was recovered.

MS m/z (ESI): 348.2 [M+1]

Step 5 Preparation of5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide p-toluenesulfonate

5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 11d (20 mg, 0.057 mmol) and p-toluenesulfonic acidmonohydrate (12 mg, 0.063 mmol) were dissolved in 2 mL ofdichloromethane with stirring upon cooling by an ice-water bath. Thereaction mixture was stirred for 10 minutes at the same temperature andthe ice-water bath was removed. The mixture was concentrated underreduced pressure to obtain an oil. The mixture was treated with 5 mL ofethyl acetate with stirring to produce a white precipitate. The mixturewas filtered under reduced pressure to obtain the title compound5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide p-toluenesulfonate 11 (0.026 g, yield 86.8%) as awhite solid.

MS m/z (ESI): 348.2 [M+1]

¹H NMR (CDCl₃, 400 MHz): δ 7.68 (d, 2H), 7.17 (d, 2H), 3.72 (m, 1H),3.68 (m, 1H), 3.58 (m, 1H), 3.30 (m, 4H), 2.98 (s, 2H), 2.81 (s, 6H),2.55 (m, 2H), 2.22 (m, 4H), 2.05 (m, 2H), 1.62 (m, 2H), 1.56 (s, 3H).

Example 125-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide p-toluenesulfonate

Step 1 Preparation of5-cyano-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acid dimethylamide

N,N-dimethyl-5-oxo-hexahydro-cyclopenta[c]pyrrole-2-carboxamide 1g (12.9g, 0.066 mol) and 4-toluenesulfonylmethyl isocyanide (14.2 g, 0.0727mol) were dissolved in 240 mL of 1,2-dimethoxyethane with stirring uponcooling by an ice-water bath, followed by dropwise addition of asolution of potassium tert-butoxide (14.8 g, 0.132 mol) in tent-butanol.Upon completion of the addition, the ice-water bath was removed. Thereaction mixture was warmed up to room temperature and reactedovernight. The reaction was monitored by TLC until the disappearance ofthe starting materials. The mixture was diluted with 100 mL of water and50 mL of saturated brine and extracted with ethyl acetate (200 mL×3).The combined organic extracts were dried over anhydrous magnesiumsulfate, filtered and concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography to obtain thetitle compound 5-cyano-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 12a (7.0 g, yield 51%) as a light yellow oil.

MS m/z (ESI): 208.1 [M+1]

¹H NMR (DMSO-D₆, 400 MHz): δ 3.5-3.0 (m, 4H), 2.75 (s, 6H), 2.6 (m, 1H),2.1-1.5 (m, 6H).

Step 2 5-Cyano-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

5-Cyano-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acid dimethylamide12a (4.2 g, 20.2 mmol) and iodomethane (11.5 g, 80.8 mmol) weredissolved in 100 mL of tetrahydrofuran followed by dropwise addition oflithium hexamethyldisilazide (80.8 mL, 80.8 mmol) at room temperatureunder nitrogen atmosphere. The reaction mixture was reacted for 2 hoursuntil MS showed the disappearance of the starting materials. The mixturewas diluted with 100 mL of water and extracted with ethyl acetate (200mL×3). The combined organic extracts were dried over anhydrous sodiumsulfate, filtered and concentrated under reduced pressure to obtain anoil. TLC showed that there are two adjacent spots, and then the oil waspurified by silica gel column chromatography to obtain the bottom spotcompound 5-cyano-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 12b (2.411 g) as a light yellow oil.

MS m/z (ESI): 222.2 [M+1]

¹H NMR (CDCl₃, 400 MHz): δ 3.44 (m, 2H), 3.3 (m, 2H), 2.85 (s, 6H), 2.77(m, 2H), 2.06 (m, 2H), 2.00 (m, 2H), 1.36 (s, 3H).

Step 3 Preparation of2-dimethylcarbamoyl-5-methyl-octahydro-cyclopenta[c]pyrrole-5-carboxylicacid

5-Cyano-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 12b (6.99 g, 31.6 mmol) was dissolved in 90 mL ofhydrochloric acid (36%). The reaction mixture was heated at 50° C. in anoil bath and stirred for 48 hours. The reaction was monitored by TLCuntil the disappearance of the starting materials. The reaction mixturewas diluted with 100 mL of water. Upon cooling by an ice-water bath, themixture was adjusted to pH 6 with potassium carbonate and extracted withethyl acetate (1500 mL×3) and dichloromethane (150 mL×3) successively.The combined organic extracts were dried over anhydrous sodium sulfate,filtered and concentrated under reduced pressure to obtain the titlecompound2-dimethylcarbamoyl-5-methyl-octahydro-cyclopenta[c]pyrrole-5-carboxylicacid 12c (7.0 g, yield 92%) as a yellow liquid, which was directly usedin the next step.

MS m/z (ESI): 241.2 [M+1]

Step 4 Preparation of2-dimethylcarbamoyl-5-methyl-hexahydro-cyclopenta[c]pyrrole-5-carbonylazide

Upon cooling by an ice-water bath,2-dimethylcarbamoyl-5-methyl-octahydro-cyclopenta[c]pyrrole-5-carboxylicacid 12c (1.0 g, 4.2 mmol) was dissolved in 25 mL of acetone withstirring, followed by dropwise addition of a solution of triethylamine(463.5 mg, 4.58 mmol) and ethyl chloroformate (497 mg, 4.58 mmol) in 10mL of acetone at −5° C. The reaction mixture was reacted for 15 minutesat −5° C. followed by addition of a solution of sodium azide (546 mg,8.4 mmol) in 10 mL of water. Then the reaction mixture was stirred foranother 30 minutes at −5° C., and quenched with 25 mL of water andextracted with ethyl acetate (50 mL×3). The combined organic extractswere dried over anhydrous sodium sulfate, filtered and concentratedunder reduced pressure to obtain the crude compound2-dimethylcarbamoyl-5-methyl-hexahydro-cyclopenta[c]pyrrole-5-carbonylazide12d (700 mg) as a yellow oil.

Step 5 to Step 6

2-Dimethylcarbamoyl-5-methyl-hexahydro-cyclopenta[c]pyrrole-5-carbonylazide12d (700 mg) was added to 20 mL of toluene and the reaction mixture washeated to reflux for 2 hours. The toluene was evaporated resulting inthe formation of(3aS,5r,6aR)-5-isocyanato-5-methyl-N,N,5-trimethyl-hexahydro-cyclopenta[c]pyrrole-2(1H)-2-carboxamide12e. Upon cooling by an ice-water bath, to a solution of 8 mL ofhydrochloric acid (8 N) was added the above mentioned mixture. Thereaction mixture was stirred for 30 minutes and the reaction wasmonitored by TLC until the disappearance of the starting materials. Uponcooling by an ice-water bath, the mixture was adjusted to pH>12 with a 3N aqueous sodium hydroxide solution and extracted with ethyl acetate (15mL×3). The combined organic extracts were dried over anhydrous magnesiumsulfate, filtered and concentrated under reduced pressure. The residuewas purified by silica gel column chromatography to obtain the titlecompound 5-amino-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 12f (500 mg) as a light yellow oil.

MS m/z (ESI): 212.2 [M+1]

¹H NMR (DMSO-D₆, 400 MHz): δ 3.5-3.0 (m, 4H), 2.72 (s, 6H), 1.7-1.3 (m,6H), 1.04 (s, 3H).

Step 7 Preparation of5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

5-Amino-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 12f (332 mg, 1.57 mmol) was dissolved in 10 mL of asolvent mixture of N,N-dimethylformamide/dichloromethane (1:1) followedby addition of 1-(2-chloro-acetyl)-pyrrolidine-2-cyano (217 mg, 1.26mmol). The reaction mixture was reacted overnight at room temperatureand concentrated under reduced pressure. The resulting residue waspurified by silica gel column chromatography to obtain the titlecompound5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 12g (240 mg, yield 55%) as a colorless oil.

MS m/z (ESI): 348.2 [M+1]

Step 8 Preparation of5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide p-toluenesulfonate

5-[2-(2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopnta[c]pyrrole-2-carboxylicacid dimethylamide 12g (150 mg, 0.43 mmol) and p-toluenesulfonic acidmonohydrate (82 mg, 0.43 mmol) were dissolved in 4 mL of dichloromethanewith stirring. The reaction mixture was stirred for 10 minutes andconcentrated under reduced pressure to obtain the title compound5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide p-toluenesulfonate 12 (0.022 g, yield 95.3%) as alight yellow solid.

MS m/z (ESI): 348.2 [M+1]

¹H NMR (DMSO-D₆, 400 MHz): δ 4.82 (m, 1H), 3.97 (s, 2H), 3.79 (m, 2H),3.49 (m, 2H), 3.21 (m, 4H), 2.75 (s, 6H), 2.62 (m, 2H), 2.19 (m, 2H),1.92-1.61 (m, 3H), 2.06 (m, 3H), 1.2 (s, 3H).

Example 135-[2-((2S,4S)-2-Cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamidetartrate

Step 1 Preparation of(2S,4R)-1-(tert-butoxycarbonyl)-4-hydroxypyrrolidine-2-carboxylic acid

L-Hydroxyproline 13a (60 g, 0.458 mol) was added to a solution of 750 mLof solvent mixture of tetrahydrofuran/water (2:1) followed by 252 mL ofaqueous sodium hydroxide (10%) and a solution of di-tent-butyldicarbonate (136 g, 0.624 mol) in 750 mL of solvent mixture oftetrahydrofuran/water (2:1). The reaction mixture was reacted overnightat room temperature and diluted with 500 mL of ethyl acetate. The layerswere separated and then the organic layer was discarded, and the aqueouslayer was adjusted to pH 2 with concentrated hydrochloric acid. Themixture was extracted with 1.5 L of ethyl acetate. The combined organicextracts were washed with saturated brine, dried over anhydrous sodiumsulfate, filtered and concentrated under reduced pressure to obtain thetitle compound(2S,4R)-1-(tert-butoxycarbonyl)-4-hydroxypyrrolidine-2-carboxylic acid13b (86.4 g, yield 80%) as a colorless oil.

Step 2 Preparation of (2S,4R)-tent-butyl2-carbamoyl-4-hydroxypyrrolidine-1-carboxylate

(2S,4R)-1-(tert-Butoxycarbonyl)-4-hydroxypyrrolidine-2-carboxylic acid13b (86.4 g, 0.374 mol) was dissolved in 1.2 L of tetrahydrofuranfollowed by addition of triethylamine (41 g, 0.411 mol) under argonatmosphere. The reaction mixture was cooled to −15° C., ethylchlorformate (43.84 g, 0.411 mol) was added. After the mixture wasstirred for 10 minutes, aqueous ammonia (236.8 mL) was added. Thereaction mixture was slowly warmed up to 5° C. during 2 hours followedby addition of ammonium chloride (32 g). The reaction mixture wasstirred for 30 minutes and separated. The separated organic phase wasdried over anhydrous sodium sulfate, and the aqueous phase was extractedwith ethyl acetate (100 mL×2). The combined organic extracts wereconcentrated under reduced pressure to obtain the title compound(2S,4R)-tert-butyl 2-carbamoyl-4-hydroxypyrrolidine-1-carboxylate 13c(74 g, yield 86%) as a white solid.

Step 3 Preparation of (2S,4R)-tert-butyl2-cyano-4-hydroxypyrrolidine-1-carboxylate

(2S,4R)-tert-Butyl 2-carbamoyl-4-hydroxypyrrolidine-1-carboxylate 13c(74 g, 0.3217 mol) was dissolved in 740 mL of pyridine with stirringunder argon atmosphere. The mixture was cooled to −20° C. followed bydropwise addition of trifluoroacetic anhydride (169 g, 0.804 mol). Uponcompletion of the addition, the reaction mixture was warmed up to roomtemperature. The reaction was monitored by TLC until the disappearanceof the starting materials. The mixture was quenched with water anddiluted with 0.8 L of ethyl acetate. The separated organic phase waswashed with 500 mL of saturated brine and neutralized with 400 mL ofconcentrated hydrochloric acid to slight acidity. The resulting mixturewas washed with 300 mL of aqueous sodium hydroxide solution (2 M) and500 mL saturated brine successively, dried over anhydrous sodiumsulfate, filtered and concentrated under reduced pressure to obtain thetitle compound (2S,4R)-tert-butyl2-cyano-4-hydroxypyrrolidine-1-carboxylate 13d (49.7 g, yield 73%) as abrown oil.

Step 4 Preparation of (2S,4S)-tert-butyl2-cyano-4-fluoropyrrolidine-1-carboxylate

(2S,4R)-tert-Butyl 2-cyano-4-hydroxypyrrolidine-1-carboxylate 13d (49.7g, 0.2344 mol) was dissolved in 1130 mL of dichloromethane with stirringunder argon atmosphere. The mixture was cooled to −30° C. followed byaddition of diethylaminosulfur trifluoride (56.7 g, 0.3516 mol). Afterstirred for 45 minutes, the reaction mixture was warmed up to −5° C.Then the mixture was reacted overnight at room temperature. The reactionwas monitored by TLC until the disappearance of the starting materials.The reaction mixture was neutralized with saturated aqueous sodiumcarbonate solution to pH >7 at such a rate that the reaction temperaturewas kept below 20° C. Then ice water and 500 mL of dichloromethane wereadded. The separated organic phase was washed with 500 mL of saturatedaqueous sodium hydrogen sulfate and 500 mL of saturated brinesuccessively, dried over anhydrous sodium sulfate, filtered andconcentrated under reduced pressure in which the concentrationtemperature was kept below 38° C. to obtain the title compound(2S,4S)-tent-butyl 2-cyano-4-fluoropyrrolidine-1-carboxylate 13e (50 g,yield 100%) as a light yellow solid.

Step 5 Preparation of (2S,4S)-4-fluoropyrrolidine-2-carbonitrile

(2S,4S)-tert-Butyl 2-cyano-4-fluoropyrrolidine-1-carboxylate 13e (1 g,4.6 mmol) was dissolved in 2 mL of ethyl acetate with stirring underargon atmosphere. The mixture was cooled to 15° C. followed by additionof a solution of hydrogen chloride in 2.5 mL of 1,4-dioxane (3 M). Thereaction mixture was stirred at room temperature for 2 hours. Then themixture was reacted for another 5 hours at 25° C. The reaction wasmonitored by TLC, which indicated the presence of the startingmaterials. The reaction mixture was filtered and the filtrate wasstirred at room temperature for 2 hours resulting in the formation of awhite precipitate. The mixture was filtered again and the filtrate wasstirred for another 2 hours, then filtered. The filter cakes werecombined to obtain the title compound(2S,4S)-4-fluoropyrrolidine-2-carbonitrile 13f (15.6 g, yield 76.6%) asa white solid.

Step 6 Preparation of(2S,4S)-1-(2-chloroacetyl)-4-fluoropyrrolidine-2-carbonitrile

Chloroacetyl chloride (11.13 g, 98.5 mmol) was dissolved in 120 mL ofdichloromethane with stirring under argon atmosphere. Upon cooling to 0°C., (2S,4S)-4-fluoropyrrolidine-2-carbonitrile 13f (11.4 g, 75.7 mmol)was dissolved in 400 mL of dichloromethane followed by addition oftriethylamine (16.1 g, 158.97 mmol). To a solution of chloroacetylchloride in dichloromethane was added the above mentioned mixture in 30minutes. The reaction mixture was reacted at 0° C. for 2 hours anddiluted with 200 mL of ice water and 150 mL of dichloromethane. Theseparated organic phase was neutralized with saturated sodium bisulfate,washed with 300 mL of water and 300 mL of saturated brine successively,dried over anhydrous sodium sulfate, filtered and concentrated underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography to obtain the title compound(2S,4S)-1-(2-chloroacetyl)-4-fluoropyrrolidine-2-carbonitrile 13g (8 g,yield 60%) as a white crystal.

Reference: WO2003002553.

Step 7 Preparation of5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-hexahydro-cyclopenta[c]pyrrole-2(1H)-carboxamide

5-Amino-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 12f (130 mg, 0.616 mmol),(2S,4S)-1-(2-chloroacetyl)-4-fluoropyrrolidine-2-carbonitrile 13g (117.4mg, 0.616 mmol) and potassium carbonate (85 mg, 0.616 mmol) weredissolved in the solvent mixture of 2 mL of dichloromethane and 2 mL ofN,N-dimethylformamide with stirring under nitrogen atmosphere. Thereaction mixture was reacted overnight at room temperature. The reactionwas monitored by TLC until the disappearance of the starting materials.The mixture was concentrated under reduced pressure to removeN,N-dimethylformamide and dichloromethane. The resulting residue waspurified by silica gel column chromatography to obtain5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-hexahydro-cyclopenta[c]pyrrole-2(1H)-carboxamide13h (0.13 g, yield 58%) as a colorless oil.

Step 8 Preparation of5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamidetartrate

5-[2-((2S,4S)-2-Cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-hexahydro-cyclopenta[c]pyrrole-2(1H)-2-carboxamide13h (0.16 g, 0.44 mmol) was dissolved in 5 mL of dichloromethane withstirring followed by dropwise addition of a solution of 5 mL of tartaricacid (65.6 mg, 0.44 mmol) in acetone. The reaction mixture was reactedfor 30 minutes resulting in the formation of a white precipitate. Themixture was filtered and the filter cake was washed with acetone toobtain the title compound5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamidetartrate 13 (0.18 g, yield 82%) as a white solid.

¹H NMR (DMSO-D₆, 400 MHz): δ 5.76 (m, 1H), 5.46 (m, 1H), 5.0 (m, 1H),4.08-4.05 (m, 4H), 3.97 (m, 2H), 3.69 (m, 2H), 2.73 (s, 6H), 2.61 (m,2H), 1.87 (m, 3H), 1.57 (m, 3H), 1.18 (s, 3H), 1.9 (m, 2H).

Example 145-Benzyl-5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate

Step 1 Preparation of5-benzyl-5-cyano-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

5-Cyano-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acid dimethylamide12a (4.0 g, 19.3 mmol) was dissolved in 60 mL of tetrahydrofuran withstirring followed by addition of benzyl chloride (5.4 g, 42.5 mmol) andlithium hexamethyldisilazide (42.5 mL, 42.5 mmol) under nitrogenatmosphere. The reaction mixture was stirred at room temperature for 4hours. The reaction was monitored by TLC until the disappearance of thestarting materials and quenched with 50 mL of water. The mixture wasextracted with ethyl acetate. The combined organic extracts were driedover anhydrous sodium sulfate, filtered and concentrated under reducedpressure. The resulting residue was purified by silica gel columnchromatography to obtain the title compound5-benzyl-5-cyano-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 14a (2.6 g, yield 46%) as a light yellow solid.

Step 2 Preparation of5-benzyl-5-formyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

5-Benzyl-5-cyano-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 14a (2.0 g, 6.7 mmol) was dissolved in 100 mL ofdichloromethane with stirring. Upon cooling to 0° C.,diisobutylaluminium hydride (20.2 mL, 20.2 mmol) was added dropwise. Thereaction was monitored by TLC until the disappearance of the startingmaterials and quenched with water. The mixture was extracted with ethylacetate. The combined organic extracts were dried over anhydrous sodiumsulfate, filtered and concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography to obtain thetitle compound5-benzyl-5-formyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 14b (729 mg, yield 36%) as a colorless oil.

Step 3 Preparation of5-benzyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carboxylicacid

5-Benzyl-5-formyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 14b (0.729 g, 2.43 mmol) was dissolved in the solventmixture of 28 mL of tetrahydrofuran and 14 mL of water under nitrogenatmosphere, followed by addition of sodium dihydrogenphosphate dihydrate(1.14 g, 7.29 mmol), sodium chlorite (0.66 g, 7.29 mmol) and2-methyl-2-butene (0.513 g, 7.32 mmol) upon cooling to 0° C. Thereaction mixture was stirred at 0° C. for 2 hours. The reaction wasmonitored by TLC until the disappearance of the starting materials. Themixture was extracted with ethyl acetate. The combined organic extractswere dried over anhydrous magnesium sulfate, filtered and concentratedunder reduced pressure. The resulting residue was purified by silica gelcolumn chromatography to obtain the title compound5-benzyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carboxylicacid 14c (0.76 g, yield 98%) as a colorless oil.

MS m/z (ESI): 317.3 [M+1]

¹H NMR (DMSO-D₆, 400 MHz): δ 7.5-7.0 (m, 5H), 3.24 (m, 2H), 3.1 (m, 2H),2.76 (s, 2H), 2.7 (s, 6H), 2.68 (m, 2H), 1.99-1.55 (m, 4H).

Step 4 Preparation of5-benzyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carbonylazide

5-Benzyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carboxylicacid 14c (0.86 g, 2.72 mmol) was dissolved in 30 mL of acetone uponcooling by an ice-water bath. When the mixture was cooled to −5° C., asolution of triethylamine (0.303 g, 2.99 mmol) and ethyl chloroformate(0.325 g, 2.99 mmol) in 15 mL of acetone were added dropwisesuccessively. The reaction mixture was stirred for 15 minutes followedby dropwise addition of a solution of sodium azide (0.353 g, 5.44 mmol)in 15 mL of water. The reaction mixture was stirred for another 30minutes. The reaction was monitored by TLC until the disappearance ofthe starting materials. The mixture was concentrated under reducedpressure and extracted with ethyl acetate. The combined organic extractswere dried over anhydrous sodium sulfate, filtered and concentratedunder reduced pressure to obtain the title compound5-benzyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carbonylazide 14d (0.9 g, yield 97%) as a light yellow oil.

Step 5 Preparation of5-benzyl-5-isocyanato-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

5-Benzyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carbonylazide 14d (1.0 g, 2.72 mmol) was dissolved in 20 mL of toluene withstirring. The reaction mixture was heated to reflux for 1.5 hours andthe solvent was evaporated to obtain the crude title compound5-benzyl-5-isocyanato-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 14e which was directly used in the next step.

Step 6 Preparation of5-amino-5-benzyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

To a solution of 12 mL hydrochloric acid (8 N) was added dropwise5-benzyl-5-isocyanato-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 14e obtained from the above step at room temperature. Thereaction mixture was stirred for 30 minutes and adjusted to pH 9 with a4 N sodium hydroxide aqueous solution. The mixture was extracted withethyl acetate. The combined organic extracts were dried over anhydroussodium sulfate, filtered and concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography toobtain the title compound5-amino-5-benzyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 14f (0.55 g, yield 70%) as a colorless oil.

Step 7 Preparation of5-benzyl-5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

5-Amino-5-benzyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 14f (0.1 g, 0.35 mmol) was dissolved in 4 mL of thesolvent mixture of dichloromethane/N,N-dimethylformamide (V/V=1/1)followed by addition of(2S,4S)-1-(2-chloroacetyl)-4-fluoropyrrolidine-2-carbonitrile 13g (66.5mg, 0.35 mmol) and potassium carbonate (49 mg, 0.35 mmol). The reactionmixture was reacted at 40° C. for 12 hours. The reaction was monitoredby TLC until the disappearance of the starting materials. The mixturewas concentrated, and the resulting residue was purified by silica gelcolumn chromatography to obtain the title compound5-benzyl-5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 14g (87 mg, yield 56%) as a white solid.

MS m/z (ESI): 442.2 [M+1]

Step 8 Preparation of5-benzyl-5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate

5-Benzyl-5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 14g (87 mg, 0.197 mmol) was dissolved in 3 mL ofdichloromethane with stirring followed by dropwise addition of asolution of 3 mL of tartaric acid in acetone. The reaction mixture wasreacted at room temperature for 30 minutes and concentrated. Theresulting residue was recrystallized from a solvent mixture of ethylacetate/acetone to obtain the title compound5-benzyl-5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate 14 (80 mg, yield 68%) as a white solid.

¹H NMR (DMSO-D₆, 400 MHz): δ 7.32-7.2 (m, 5H), 5.55 (d, 1H), 5.41 (d,1H), 4.97 (m, 1H), 4.31 (s, 2H), 4.08 (m, 1H), 4.0-3.5 (m, 6H), 2.81 (s,2H), 2.78 (s, 6H), 2.5 (m, 2H), 2.0 (m, 3H), 1.6-1.3 (m, 6H).

Example 155-Cyclohexylmethyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate

Step 1 Preparation of5-cyano-5-cyclohexylmethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

5-Cyano-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acid dimethylamide12a (3.66 g, 17.6 mmol) was dissolved in 150 mL of tetrahydrofuran withstirring followed by dropwise addition of cyclohexylmethyl bromide (6.2g, 35.2 mmol) and lithium hexamethyldisilazide (35.2 mL, 32.5 mmol). Thereaction mixture was reacted at room temperature for 2 hours. Thereaction was monitored by TLC until the disappearance of the startingmaterials. Then 150 mL of a saturated aqueous solution of ammoniumchloride was added. The mixture was extracted with ethyl acetate (60mL×3). The combined organic extracts were dried over anhydrous magnesiumsulfate, filtered and concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography to obtain thetitle compound5-cyano-5-cyclohexylmethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 15a (2.2 g, yield 41.5%) as a yellow solid.

MS m/z (ESI): 304.5 [M+1]

Step 2 Preparation of5-cyclohexylmethyl-5-formyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

Upon cooling to 0° C. by an ice-water bath,5-cyano-5-cyclohexylmethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 15a (1.2 g, 3.95 mmol) was dissolved in 30 mL ofdichloromethane with stirring followed by dropwise addition ofdiisobutylaluminium hydride (11.8 mL, 11.8 mmol). The reaction mixturewas reacted for 1 hour. The reaction was monitored by TLC until thedisappearance of the starting materials. Then a solution of saturatedaqueous potassium sodium tartrate was added. The mixture was stirreduntil the solution was clear and extracted with ethyl acetate (100mL×3). The combined organic extracts were dried over anhydrous magnesiumsulfate, filtered and concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography to obtain thetitle compound5-cyclohexylmethyl-5-formyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 15b (0.4 g, yield 33%) as a light yellow oil.

MS m/z (ESI): 307.4 [M+1]

Step 3 Preparation of5-cyclohexylmethyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carboxylicacid

5-Cyclohexylmethyl-5-formyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 15b (0.713 g, 2.33 mmol) was dissolved in the solventmixture of 60 mL of tetrahydrofuran and 30 mL of water. Upon cooling to0° C., sodium dihydrogenphosphate dihydrate (1.09 g, 6.99 mmol), sodiumchlorite (0.79 g, 6.99 mmol) and 2-methyl-2-butene (0.62 mL, 7.0 mmol)were added. The reaction mixture was reacted at 0° C. for 2 hours. Thereaction was monitored by TLC until the disappearance of the startingmaterials. The mixture was concentrated to remove tetrahydrofuran andextracted with ethyl acetate. The combined organic extracts were driedover anhydrous magnesium sulfate, filtered and concentrated underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography to obtain the title compound5-cyclohexylmethyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carboxylicacid 15c (0.75 g, yield 100%) as a yellow solid.

MS m/z (ESI): 323.3 [M+1]

Step 4 Preparation of5-cyclohexylmethyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carbonylazide

Upon cooling by an ice-water bath,5-cyclohexylmethyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carboxylicacid 15c (0.75 g, 2.33 mmol) was dissolved in 20 mL of acetone withstirring followed by dropwise addition of triethylamine (0.36 mL, 2.56mmol) and a solution of 2 mL of ethyl chloroformate (0.25 mL, 2.56 mmol)in acetone. The reaction mixture was stirred for 15 minutes and asolution of 2 mL of sodium azide (0.303 g, 4.66 mmol) in water wasadded. The reaction mixture was reacted for another 30 minutes at 0°C.˜5° C. The reaction was monitored by TLC until the disappearance ofthe starting materials. The mixture was concentrated to remove acetoneand diluted with 10 mL of water. The mixture was extracted with ethylacetate (10 mL×5). The combined organic extracts were dried overanhydrous magnesium sulfate, filtered and concentrated under reducedpressure to obtain the title compound5-cyclohexylmethyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carbonylazide 15d (0.79 g) as a yellow oil which was directly used in the nextstep.

Step 5 Preparation of5-cyclohexylmethyl-5-isocyanato-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

5-Cyclohexylmethyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carbonylazide 15d (0.79 g, 2.27 mmol) was dissolved in 20 mL of toluene and thereaction mixture was heated to reflux for 1 hour. The reaction wasmonitored by TLC until the disappearance of the starting materials. Themixture was concentrated to obtain the title compound5-cyclohexylmethyl-5-isocyanato-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 15e (0.65 g) as a grey solid, which was directly usedin the next step.

MS m/z (ESI): 320.4 [M+1]

Step 6 Preparation of5-amino-5-cyclohexylmethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

Upon cooling by an ice-water bath, to a solution of 10 mL ofhydrochloric acid (8 N) was added dropwise5-cyclohexylmethyl-5-isocyanato-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 15e (0.65 g, 2.03 mmol). Then the ice-water bath wasremoved. The reaction mixture was reacted at 50° C. for 20 minutes. Thereaction was monitored by TLC until the disappearance of the startingmaterials. The mixture was adjusted to pH>8 with concentrated ammoniaand extracted with ethyl acetate (30 mL×5). The combined organicextracts were dried over anhydrous magnesium sulfate, filtered andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography to obtain5-amino-5-cyclohexylmethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 15f (0.5 g, yield 84%) as a grey oil.

MS m/z (ESI): 294.3 [M+1]

Step 7 Preparation of5-(cyclohexylmethyl)-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-cyclohexylmethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

5-Amino-5-cyclohexylmethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 15f (0.115 g, 0.39 mmol) was dissolved in 10 mL of asolvent mixture dichloromethane/N,N-dimethylformamide (V/V=1/1) withstirring, followed by addition of1-(2-chloro-acetyl)-pyrrolidine-2-cyano (54 mg, 0.31 mmol). The reactionmixture was reacted at 50° C. for 2 hours. The reaction was monitored byTLC until the disappearance of the starting materials. The mixture wasconcentrated under reduced pressure to remove dichloromethane andN,N-dimethylformamide. The resulting residue was purified by silica gelcolumn chromatography to obtain the title compound5-(cyclohexylmethyl)-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-cyclohexylmethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 15 g (113 mg, yield 67%) as a colorless oil.

MS m/z (ESI): 430.5 [M+1].

Step 8 Preparation of5-(cyclohexylmethyl)-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate

5-(Cyclohexylmethyl)-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-cyclohexylmethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 15g (113 mg, 0.263 mmol) was dissolved in 10 mL ofethyl acetate followed by dropwise addition of a solution of 2 mL oftartaric acid in acetone. The reaction mixture was stirred for 30minutes and filtered to obtain the title compound5-(cyclohexylmethyl)-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate 15 (40 mg) as a white solid.

¹H NMR (DMSO-D₆, 400 MHz): δ 5.2 (m, 1H), 4.77 (s, 2H), 4.18 (m, 3H),4.02 (m, 1H), 3.63 (m, 2H), 3.32 (m, 2H), 2.73 (s, 6H), 2.58 (m, 2H),2.16-1.87 (m, 9H), 1.59-1.19 (m, 8H).

Example 165-Cyclopentyl-5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate

Step 1 Preparation of5-cyano-5-cyclopentyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

5-Cyano-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acid dimethylamide12a was dissolved in 30 mL of N,N-dimethylformamide followed by additionof iodo-cyclopentane (5.4 g, 27.5 mmol) and lithium hexamethyldisilazide(27.5 mL, 27.5 mmol) under nitrogen atmosphere. The reaction mixture wasstirred at room temperature for 2 hours. The reaction was monitored byTLC until the disappearance of the starting materials. The mixture wasdiluted with 10 mL of water and concentrated to removeN,N-dimethylformamide and extracted with ethyl acetate. The combinedorganic extracts were dried over anhydrous sodium sulfate, filtered andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography to obtain the title compound5-cyano-5-cyclopentyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 16a (2.6 g, yield 42%) as a light yellow solid.

MS m/z (ESI): 276.2 [M+1]

¹H NMR (DMSO-D₆, 400 MHz): δ 3.35-3.0 (m, 4H), 2.24 (s, 6H), 1.34-2.5(m, 15H).

Step 2 Preparation of5-cyclopentyl-5-formyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

5-Cyano-5-cyclopentyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 16a (0.817 g, 2.97 mmol) was dissolved in 40 mL ofdichloromethane with stirring at 0° C., followed by dropwise addition ofdiisobutylaluminium hydride (8.9 mL, 8.9 mmol). The reaction mixture wasstirred for 45 minutes. The reaction was monitored by TLC until thedisappearance of the starting materials. The mixture was concentrated toremove dichloromethane and the residue was diluted with 100 mL of asaturated aqueous potassium sodium tartrate solution. The mixture wasstirred until the solution was clear and extracted with ethyl acetate(100 mL×4). The combined organic extracts were dried over anhydrousmagnesium sulfate, filtered and concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography toobtain the title compound5-cyclopentyl-5-formyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 16b (0.266 g, yield 32%) as a light yellow oil.

MS m/z (ESI): 279.3 [M+1]

Step 3 Preparation of5-cyclopentyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carboxylicacid

5-Cyclopentyl-5-formyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 16b (0.266 g, 0.955 mmol) was dissolved in a solventmixture of 20 mL of tetrahydrofuran and 10 mL of water. Then sodiumdihydrogenphosphate dihydrate (0.448 g, 2.87 mmol), sodium chlorite(0.26 g, 2.87 mmol) and 2-methyl-2-butene (0.24 mL, 2.88 mmol) wereadded at 0° C. The reaction mixture was stirred at 0° C. for 2 hours.The reaction was monitored by TLC until the disappearance of thestarting materials. The mixture was concentrated to removetetrahydrofuran and extracted with ethyl acetate. The combined organicextracts were dried over anhydrous magnesium sulfate, filtered andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography to obtain the title compound5-cyclopentyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carboxylicacid 16c (0.28 g, yield 99.6%) as a yellow solid.

MS m/z (ESI): 295.5 [M+1]

Step 4 Preparation of5-cyclopentyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carbonylazide

5-Cyclopentyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carboxylicacid 16c (0.28 g, 6.95 mmol) was dissolved in 20 mL of acetone uponcooling by an ice-water bath, followed by addition of triethylamine(0.15 mL, 1.05 mmol) and a solution of 2 mL of ethyl chloroformate (0.1mL, 1.05 mmol) in acetone at 0° C.˜−5° C. After the reaction mixture wasstirred for 15 minutes, a solution of sodium azide (0.124 g, 1.9 mmol)in water was added dropwise. The reaction mixture was stirred foranother 30 minutes. The reaction was monitored by TLC until thedisappearance of the starting materials. The mixture was concentratedand the residue was diluted with 10 mL of water. The mixture wasextracted with ethyl acetate. The combined organic extracts were driedover anhydrous magnesium sulfate, filtered and concentrated underreduced pressure to obtain the title compound5-cyclopentyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carbonylazide 16d (0.287 g, yield 95%) as a yellow oil which was directly usedin the next step.

Step 5 Preparation of5-cyclopentyl-5-isocyanato-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

5-Cyclopentyl-2-dimethylcarbamoyl-octahydro-cyclopenta[c]pyrrole-5-carbonylazide 16d (0.287 g, 0.9 mmol) was dissolved in 10 mL of toluene withstirring. The reaction mixture was heated to reflux for 1 hour. Thereaction was monitored by TLC until the disappearance of the startingmaterials. The mixture was concentrated to obtain the title compound5-cyclopentyl-5-isocyanato-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 16e which was directly used in the next step.

MS m/z (ESI): 292.3 [M+1]

Step 6 Preparation of5-amino-5-cyclopentyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

Upon cooling by an ice-water bath, to a solution of 10 mL hydrochloricacid (8 N) was added dropwise5-cyclopentyl-5-isocyanato-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 16e obtained from the above step. Then the ice-waterbath was removed and the reaction mixture was reacted at 50° C. for 15minutes. The reaction was monitored by TLC until the disappearance ofthe starting materials. The mixture was adjusted to pH>8 withconcentrated ammonia and extracted with ethyl acetate. The combinedorganic extracts were dried over anhydrous magnesium sulfate, filteredand concentrated under reduced pressure. The resulting residue waspurified by silica gel column chromatography to obtain the titlecompound5-amino-5-cyclopentyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 16f (0.18 g, yield 81.8%) as a yellow oil.

MS m/z (ESI): 266.2 [M+1]

Step 7 Preparation of5-cyclopentyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

5-Amino-5-cyclopentyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 16f (0.108 g, 0.407 mmol) and1-(2-chloro-acetyl)-pyrrolidine-2-cyano (70 mg, 0.407 mmol) weredissolved in 3 mL of N,N-dimethylformamide with stirring followed byaddition of potassium carbonate (57 mg, 0.407 mmol) under nitrogenatmosphere. The reaction mixture was reacted overnight at 80° C. uponheating by an oil bath. The reaction was monitored by TLC until thedisappearance of the starting materials. The mixture was concentrated,and the resulting residue was purified by silica gel columnchromatography to obtain the title compound5-cyclopentyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 16g (100 mg, yield 61.3%) as a colorless oil.

MS m/z (ESI): 402.3 [M+1]

Step 8 Preparation of5-cyclopentyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate

5-Cyclopentyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 16g (102 mg, 0.254 mmol) was dissolved in 2 mL ofethyl acetate followed by addition of a solution of 3 mL of tartaricacid in acetone. The reaction mixture was stirred for 30 minutesresulting in the formation of a white precipitate followed by additionof n-hexane, and stirred again. The mixture was filtered to obtain thetitle compound5-cyclopentyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate 16 (88 mg, yield 65%) as a white solid.

Example 175-Benzyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate

Step 1 Preparation of5-benzyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

5-Amino-5-benzyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 14f (0.1 g, 0.35 mmol),1-(2-chloro-acetyl)-pyrrolidine-2-cyano (120.4 mg, 0.7 mmol) andpotassium carbonate (49 mg, 0.35 mmol) were dissolved in 3 mL ofN,N-dimethylformamide under nitrogen atmosphere. The reaction mixturewas reacted overnight at 80° C. upon heating by an oil bath. Thereaction was monitored by TLC until the disappearance of the startingmaterials. The mixture was concentrated, and the resulting residue waspurified by silica gel column chromatography to obtain the titlecompound5-benzyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 17a (56 mg, yield 40%) as a colorless oil.

MS m/z (ESI): 424.3 [M+1]

Step 2 Preparation of5-benzyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate

5-Benzyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 17a (56 mg, 0.132 mmol) was dissolved in 1 mL ofethyl acetate with stirring followed by addition of a solution of 1 mLof tartaric acid in acetone. The reaction mixture was stirred for 30minutes resulting in the formation of a white precipitate followed byaddition of n-hexane, and stirred again. The mixture was filtered toobtain the title compound5-benzyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate 17 (50 mg, yield 67%) as a white solid.

¹H NMR (DMSO-D₆, 400 MHz): δ 7.32-7.19 (m, 5H), 4.78 (m, 1H), 4.23 (s,2H), 3.8-3.0 (m, 8H), 2.75 (s, 2H), 2.73 (s, 6H), 2.18 (m, 2H), 2.16 (m,2H), 1.87 (m, 3H), 1.35 (m, 3H).

Example 185-[2-((2S,4S)-2-Cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide p-toluenesulfonate

Step 1 Preparation of5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

5-Amino-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 11c (1.3 g, 7.58 mmol),(2S,4S)-1-(2-chloroacetyl)-4-fluoropyrrolidine-2-carbonitrile 13g (1.74g, 9.1 mmol), potassium carbonate (1.26 g, 9.1 mmol), 30 mL ofN,N-dimethylformamide and 18 mL of dichloromethane were added to a flaskunder nitrogen atmosphere. The reaction mixture was reacted forovernight at 30° C. upon heating by an oil bath. The reaction wasmonitored by TLC until the disappearance of the starting materials. Themixture was concentrated to remove N,N-dimethylformamide. The resultingresidue was purified by silica gel column chromatography to obtain thetitle compound5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 18a (1.39 g, yield 50%) as a white solid.

MS m/z (ESI): 348.2 [M+1]

Step 2 Preparation of5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide p-toluenesulfonate

5-[2-(2-Cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 18a (0.9 g, 2.47 mmol) was dissolved in 5 mL ofdichloromethane with stirring at room temperature. To a solution ofp-toluenesulfonic acid monohydrate (469 mg, 2.47 mmol) in 3 mL ofacetone was added the above mentioned solution, which resulted in theformation of a white precipitate. The reaction mixture was stirred for30 minutes followed by addition of 1 mL of n-hexane. The mixture wasfiltered to obtain the title compound5-[2-((2S,4S)-2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-methyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide p-toluenesulfonate 18 (1.3 g, yield 93%) as a whitesolid.

MS m/z (ESI): 366.1 [M+1]

¹H NMR (DMSO-D₆, 400 MHz): δ 7.74 (d, 2H), 7.29 (d, 2H), 5.57-5.44 (m,1H), 5.07 (m, 1H), 4.19-4.07 (m, 4H), 3.49 (m, 2H), 3.35 (m, 2H), 3.0(m, 2H), 2.9 (s, 6H), 2.6 (m, 2H), 2.39 (m, 5H), 1.57 (m, 2H), 1.34 (s,3H).

Example 195-Ethyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate

Step 1 5-Cyano-5-ethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

5-Cyano-hexahydro-cyclopenta[c]pyrrole-2-carboxylic acid dimethylamide12a (3.0 g, 14.5 mmol) was dissolved in 60 mL of tetrahydrofuran withstirring followed by addition of ethyl iodide (4.52 g, 29 mmol) andlithium hexamethyldisilazide (29 mL, 29 mmol) under nitrogen atmosphere.The reaction mixture was reacted at room temperature for 2 hours. Thereaction was monitored by TLC until the disappearance of the startingmaterials. The mixture was diluted with 20 mL of water and extractedwith ethyl acetate (200 mL×3). The combined organic extracts were driedover anhydrous sodium sulfate, filtered and concentrated under reducedpressure to obtain an oil. TLC showed that there are two adjacent spots,and then the oil was purified by silica gel column chromatography toobtain the title compound5-cyano-5-ethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 19a (1.64 g) as a light yellow solid.

MS m/z (ESI): 236.3 [M+1]

¹H NMR (DMSO-D₆, 400 MHz): δ 3.5-3.0 (m, 4H), 2.7 (s, 6H), 1.9-1.2 (m,8H), 1.19 (m, 3H).

Step 2 Preparation of5-ethyl-5-formyl-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamide

5-Cyano-5-ethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 19a (1.54 g, 6.55 mmol) was dissolved in 60 mL ofdichloromethane. Diisobutylaluminium hydride (19.6 mL, 19.6 mmol) wasadded dropwise at 0° C. The reaction mixture was stirred at 0° C. for 30minutes. The reaction was monitored by TLC until the disappearance ofthe starting materials. The mixture was quenched with 1.5 mL of water,dried over anhydrous magnesium sulfate, filtered and concentrated underreduced pressure to obtain the title compound5-ethyl-5-formyl-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxamide19b (0.4 g, yield 26%) as a light yellow oil.

MS m/z (ESI): 239.1 [M+1]

Step 3 Preparation of5-ethyl-2-dimethylcarbamoyl-hexahydro-cyclopenta[c]pyrrole-5-carboxylicacid

5-Ethyl-5-formyl-N,N-dimethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 19b (0.4 g, 1.68 mmol) was dissolved in the solventmixture of 18 mL of tetrahydrofuran and 9 mL of water, followed byaddition of sodium dihydrogenphosphate dihydrate (787 mg, 5.04 mmol),sodium chlorite (0.454 g, 5.04 mmol) and 2-methyl-2-butene (354 mg, 5.06mmol) at 0° C. The reaction mixture was reacted at 0° C. for 2 hoursunder nitrogen atmosphere. The reaction was monitored by TLC until thedisappearance of the starting materials. The mixture was extracted withethyl acetate. The combined organic extracts were dried over anhydrousmagnesium sulfate, filtered and concentrated under reduced pressure toobtain the title compound5-ethyl-2-dimethylcarbamoyl-hexahydro-cyclopenta[c]pyrrole-5-carboxylicacid 19c (0.426 g, yield 100%) as a yellow solid which was directly usedin the next step.

MS m/z (ESI): 255.2 [M+1]

Step 4 Preparation of5-ethyl-2-dimethylcarbamoyl-hexahydro-cyclopenta[c]pyrrole-5-carbonylazide

5-Ethyl-2-dimethylcarbamoyl-hexahydro-cyclopenta[c]pyrrole-5-carboxylicacid 19c (0.56 g, 2.2 mmol) was dissolved in 60 mL of acetone uponcooling by an ice-water bath. When cooled to −5° C., triethylamine(0.245 mg, 2.43 mmol) and a solution of 2 mL of ethyl chloroformate (263mg, 2.43 mmol) in acetone were added dropwise. After the reactionmixture was stirred for 15 minutes, a solution of sodium azide (0.286 g,4.4 mmol) in 2 mL of water was added. The reaction mixture was stirredat −5° C.˜0° C. for another 30 minutes. The reaction was monitored byTLC until the disappearance of the starting materials. The mixture wasdiluted with 10 mL of water, extracted with ethyl acetate (10 mL×5). Thecombined organic extracts were dried over anhydrous magnesium sulfate,filtered and concentrated under reduced pressure to obtain the titlecompound5-ethyl-2-dimethylcarbamoyl-hexahydro-cyclopenta[c]pyrrole-5-carbonylazide 19d (0.5 g) as a light yellow oil which was directly used in thenext step.

Step 5 Preparation of5-ethyl-5-isocyanato-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

5-Ethyl-2-dimethylcarbamoyl-hexahydrocyclopenta[c]pyrrole-5-carbonylazide 19d (0.5 g, 1.86 mmol) was dissolved in 30 mL of toluene. Thereaction mixture was heated to reflux for 2 hour. The reaction wasmonitored by TLC until the disappearance of the starting materials. Themixture was concentrated to obtain the title compound5-ethyl-5-isocyanato-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 19e, which was directly used in the next step.

Step 6 Preparation of5-amino-5-ethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide

To a solution of 12 mL hydrochloric acid (8 N) was added5-ethyl-5-isocyanato-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 19e obtained from the above step at room temperature. Thereaction mixture was stirred for 30 minutes and adjusted to pH 9-10 withan 8 N aqueous sodium hydroxide solution. The separated aqueous layerwas extracted with dichloromethane (30 mL×5). The combined organicextracts were dried over anhydrous magnesium sulfate, filtered andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography to obtain the title compound5-amino-5-ethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 19f (0.3 g, yield 71%) as a colorless oil.

MS m/z (ESI): 226.2 [M+1]

Step 7 Preparation of5-ethyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide

5-Amino-5-ethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylic aciddimethylamide 19f (0.202 g, 0.897 mmol),1-(2-chloro-acetyl)-pyrrolidine-2-cyano (185 mg, 1.07 mmol), potassiumcarbonate (148 mg, 1.07 mmol) and a solvent mixture of 9 mL ofN,N-dimethylformamide/dichloromethane (V/V=1/1) were added to a flaskunder nitrogen atmosphere. The reaction mixture was reacted at 60° C.for 2 hours. The reaction was monitored by TLC until the disappearanceof the starting materials. The mixture was concentrated under reducedpressure to remove dichloromethane and N,N-dimethylformamide. Theresulting residue was purified by silica gel column chromatography toobtain the title compound5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-ethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 19g (120 mg, yield 40%) as a light yellow oil.

MS m/z (ESI): 362.2 [M+1]

Step 8 Preparation of5-ethyl-5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate

5-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-ethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide 19g (120 mg, 0.33 mmol) was dissolved in 1 mL ofethyl acetate followed by dropwise addition of a solution of 2 mL oftartaric acid (50 mg, 0.33 mmol) in acetone. The reaction mixture wasstirred for 30 minutes and filtered to obtain the title compound5-[2-((S)-2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-5-ethyl-hexahydro-cyclopenta[c]pyrrole-2-carboxylicacid dimethylamide tartrate 19 (160 mg, yield 94%) as a white solid.

MS m/z (ESI): 362.2 [M+1]

¹H NMR (DMSO-D₆, 400 MHz): δ 4.79 (m, 1H), 4.11 (s, 2H), 3.65 (m, 2H),3.49 (m, 2H), 3.22 (m, 4H), 2.75 (s, 6H), 2.55 (m, 2H), 2.19 (m, 2H),2.09 (m, 2H), 1.99 (m, 2H), 1.51 (m, 4H), 0.86 (t, 3H).

Test Examples Biological Assays Assay for DPP-IV/DPP-VIII/DPP-IXInhibitory Activity

The following methods can be used to measure the activities of thecompounds of the present disclosure that inhibit the enzymatic activityof DPP-IV/DPP-VIII/DPP-IX. The compounds of the present disclosure aretested for their ability to inhibit the enzyme activity of purifiedDPP-IV/DPP-VIII/DPP-IX. The inhibitory rate or the half inhibitionconcentration IC₅₀ (concentration of the test compound at which 50% ofthe enzyme activity is inhibited) for each compound is determined byincubating fixed amounts of mixed enzyme substrates with severaldifferent concentrations of test compounds.

Assay for -DPP-IV Inhibitory Activity

Materials and Methods:

Materials:

a. 96-well white plate (BMG).

b. Tris buffer: in order to prepare 100 mL of 2 mM Tris Buffer, 0.0242 gTris was dissolved in about 90 mL distilled H₂O (dH₂O) firstly.Hydrochloric acid and sodium hydroxide was used to adjust pH to 8.00 andthen the solution was diluted with dH₂O to 100 mL in total volume.

c. DPP-IV enzyme (Calbiochem Catalog No. 317630) was dissolved in Trisbuffer to make a 2 mM solution.

d. DPP-IV—Glo™ substrate (Promega Catalog No. G8350) was dissolved indH₂O to make a 1 mM solution.

e. DPP-IV—Glo buffer solution (Promega Catalog No. G8350).

f. Luciferin detection reagent (Promega Catalog No. G8350).

g. DMSO.

h. dH₂O.

Procedure:

The assay was carried out as follows:

1. DPP-IV-Glo. buffer was thawed and equilibrated to room temperatureprior to use.

2. The freezing luciferin detection reagent was equilibrated to roomtemperature prior to use.

3. DPP-IV—Glo. substrate was mixed with ultrapure water by vortexingbriefly to form a 1 mM solution of the substrate.

4. The luciferin detection reagent was placed in a brown bottle withaddition of DPP-IV—Glo. buffer, and the luciferin agent was dissolvedwithin 1 minute.

5. The test compounds were dissolved in DMSO at a concentration of 50times of the final concentration.

6. 2 μL of the test compound solution with 50 times concentration wasadded to each tube, and 2 μL of DMSO was added to the negative controland blank control instead.

7. 46 μL of Tris buffer was added to each tube and 48 μL of Tris bufferwas added in the blank control.

8. 2 μL of DPP-IV enzyme was added to each negative control tube andtest tube.

9. The test tubes were shaken and centrifuged, and then the substancesof the test tube were transferred to 96-well plates.

10. The substrate and DPP-IV—Glo. were mixed in a ratio of 1:49, andthis mixture was shaken throughfully and incubated at room temperaturefor 30-60 minutes prior to use.

11. The 96-well plate was sealed by a sealing film after 50 μL of themixed solution of DPP-IV—Glo. and substrate were added to each well.

12. The substances of the 96 wells were mixing slowly by using a plateshaker at 300-500 rpm/30 s, and then the plate was incubated at roomtemperature from 30 minutes to 3 hours.

13. The luminescence value was measured.

The definition of inhibitory rate:[1−(S−B)/(N−B)]*100%

S: sample

B: blank control

N: negative control

Assay for DPP-VIII Inhibitory Activity

Materials and Methods:

Materials:

a. 96-well white plate (BMG).

b. Tris buffer: in order to prepare 100 mL 2 mM Tris buffer, 0.0242 gTris was dissolved in about 90 mL dH₂O firstly. Hydrochloric acid andsodium hydroxide were used to adjust pH to 8.00 and then the solutionwas diluted with dH₂O to 100 mL in total volume.

c. DPP-VIII enzyme (Bioscience Catalog No. 80080) was dissolved in 25 mLof Tris-HCl, pH=8.0, 100 mM NaCl, 0.05% Tween-20, 50% glycerol, and 3 mMDTT Buffer. The final concentration was 0.1 ng/100 μL assay.

d. DPP-VIII—Glo™ substrate (Promega Catalog No. G8350) was dissolved indH₂O to make a 1 mM solution.

e. DPP-VIII—Glo. buffer solution (Promega Catalog No. G8350).

f. Luciferin detection reagent (Promega Catalog No. G8350).

g. DMSO.

H. dH₂O.

Procedure:

The assay was carried out as follows:

1. DPP-VIII—Glo. buffer was thawed and equilibrated to room temperatureprior to use.

2. The freezing luciferin detection reagent was equilibrated to roomtemperature prior to use.

3. DPP-VIII—Glo. substrate was mixed with ultrapure water by vortexingbriefly to form 1 mM substrate.

4. The luciferin detection reagent was placed in a brown bottle withaddition of DPP-VIII—Glo. buffer, and the luciferin agent was dissolvedwithin 1 minute.

5. The test compounds were dissolved in DMSO to 50 times of the finalconcentration.

6. 2 μL of the test compound solution with 50 times concentration wasadded to each tube, and 2 μL of DMSO was added to the negative controland blank control instead.

7. 46 μL of Tris buffer was added to each tube and 48 μL of Tris bufferwas added in blank control.

8. 2 μL of DPP-VIII enzyme was added to each negative control tube andtest tube.

9. The tested tubes were shaken and centrifuged, and then the substancesof the solution in the test tube were transferred to 96-well plates.

10. The substrate and DPP-VIII—Glo. were mixed in a ratio of 1:49, andthis mixture was shaken throughfully and incubated at room temperaturefor 30-60 minutes prior to use.

11. The 96-well plate was sealed by a sealing film after 50 μL of themixed solution of DPP-VIII—Glo. and substrate were added to each well.

12. The substances of the 96 wells were mixing slowly by using a plateshaker at 300-500 rpm/30 s, and then the plate was incubated at roomtemperature from 30 minutes to 3 hours.

13. The luminescence value was measured.

The definition of inhibitory rate:[1−(S−B)/(N−B)]*100%

S: sample

B: blank control

N: negative control

Assay for DPP-IX Inhibitory Activity

Materials and Methods:

Materials:

a. 96-well white plate (BMG);

b. Tris buffer: in order to prepare 100 mL 2 mM Tris Buffer, 0.0242 gTris was dissolved in about 90 mL dH₂O firstly. Hydrochloric acid andsodium hydroxide were used to adjust pH to 8.00 and then the solutionwas diluted with dH₂O to 100 mL in total volume;

c. DPP-IX enzyme (Bioscience Catalog No. 80090) was dissolved in 25 mLof Tris-HCl, pH=8.0, 100 mM NaCl, 0.05% Tween-20, 50% glycerol, and 3 mMDTT Buffer. The final concentration was 0.1 ng/100 μL assay,

d. DPP-IX—Glo™ substrate (Promega Catalog No. G8350), dissolved in dH₂Oto make a 1 mM solution.

e. DPP-IX—Glo. buffer solution (Promega Catalog No. G8350).

f. Luciferin detection reagent (Promega Catalog No. G8350).

g. DMSO.

h. dH₂O.

Procedure:

1. The assay was carried out as follows:

2. DPP-IX—Glo. buffer was thawed and equilibrated to room temperatureprior to use.

3. The freezing luciferin detection reagent was equilibrated to roomtemperature prior to use.

4. DPP-IX—Glo. substrate was mixed with ultrapure water by vortexingbriefly to form 1 mM substrate.

5. The luciferin detection reagent was placed in a brown bottle withaddition of DPP-IX—Glo. buffer, and the luciferin agent was dissolvedwithin 1 minute.

6. The test compounds were dissolved in DMSO at a concentration of 50times the final concentration.

7. 2 μL of the test compound solution with 50 times concentration wasadded to each tube, and 2 μL of DMSO was added to negative control andblank control for instead.

8. 46 μL of Tris buffer was added to each tube and 48 μL of Tris bufferwas added in blank control.

9. 2 μL of DPP-1× enzyme was added to each negative control tube andtest tube.

10. The test tubes were shaken and centrifuged, and then the substancesof the test tube were transferred to 96-well plates.

11. The substrate and DPP-IX—Glo. were mixed in a ratio of 1:49, andthis mixture was shaken throughfully and incubated at room temperaturefor 30-60 minutes prior to use.

12. The 96-well plate was sealed by a sealing film after 50 μL of themixed solution of DPP-IX—Glo. and substrate were added to each well.

13. The substances of the 96 wells were mixing slowly by using a plateshaker at 300-500 rpm/30 s, and then the plate was incubated at roomtemperature from 30 minutes to 3 hours.

The luminescence value was measured.

The definition of inhibitory rate:[1−(S−B)/(N−B)]*100%

S: sample

B: blank control

N: negative control

IC₅₀ of the DPP-IV/DPP-VIII/DPP-IX of the test compounds were showed intable 1.

TABLE 1 IC₅₀ Assay Results of Examples IC50(nM) IC50(nM) IC50(nM)Example DPP-IV DPP-VIII DPP-IX 1 16 17380 5700 2 24 18240 5040 3 14 4 697 50 8 39 10 83 11 13 38480 2434 12 13 4890 16900 13 10 371 516 18 393320 380

Preliminary Evaluation of Hypoglycemic Effects of DPP-IV Inhibitors

Objective:

To observe the effects on oral glucose tolerance of the DPP-IVinhibitors the compounds of Example 1 and Example 2 in normal ICR mice,the hypoglycemic effects in vivo have been evaluated.

Test Animals:

Species, strains: ICR mice

Source: Chinese Academy of Sciences, Shanghai Laboratory Animal Center,Certification No.: SYXK (Shanghai) 2004-2005

Weight: 25-30 g

Sex: Male

Animal Number: 40

Rearing conditions: SPF-class animal room raising, temperature: 22-24°C., Humidity: 45-80%, illumination: 150-300Lx, day and night cycle withan interval of 12 hours.

Drugs:

Name: the compound of Example 1

Lot Number: 01

Color, form: white powder

Purity: 96.97%

Provided by: Shanghai Hengrui Medicine Co., Ltd.

Preparation Method: Compounds were Weighed Accurately, and thendissolved in double distilled water. The suspensions of 0.5, 0.15 and0.05 mg/mL were prepared, respectively. (Note: Although the productinstruction displayed the test compounds were soluble in water, but inthe experiment it was poorly water-soluble, i.e., at a low concentrationit can be dissolved, but at the concentration of 0.5 mg/mL, there arestill visible particles to the naked eye. 1% CMC was tried to suspendthe compounds, while it was not better than double-distilled water.)

Dose: 1, 3, 10 mg/kg by gavage. The volume is 20 mL/kg.

Name: the compound of Example 2

Lot Number: 01

Color, form: white powder

Purity: 96.62%

Provided by: Shanghai Hengrui Medicine Co., Ltd.

Preparation Method: Compounds were Weighed Accurately, and thenDissolved in double distilled water and fully mixed to prepare a 1.5mg/mL solution, and then diluted into 0.5, 0.15 and 0.05 mg/mLtransparent solutions, respectively.

Dose: 1, 3, 10 mg/kg by gavage. The volume is 20 mL/kg.

Method:

1. The effects of compounds on blood glucose levels in normal ICR mice

Normal male ICR mice were randomly grouped according to weights, 6 micein each group. The groups included a black control group as well asdifferent doses of the treatment groups as follows:

Test 1:

Blank control: double-distilled water by gavage.

Group 1: the compound of Example 1, 1 mg/kg by gavage;

the compound of Example 1, 3 mg/kg by gavage; and

the compound of Example 1, 10 mg/kg by gavage.

Group 2: the compound of Example 2, 1 mg/kg by gavage;

the compound of Example 2, 3 mg/kg by gavage; and

the compound of Example 2, 10 mg/kg by gavage.

Test 2:

Blank control: double-distilled water by gavage.

Group 1: the compound of Example 1, 1 mg/kg by gavage;

the compound of Example 1, 3 mg/kg by gavage; and

the compound of Example 1, 10 mg/kg by gavage.

Group 2: the compound of Example 2, 1 mg/kg by gavage;

the compound of Example 2, 3 mg/kg by gavage; and

the compound of Example 2, 10 mg/kg by gavage.

Animals in each group had been fasted for 6 hours, and then pretreatedwith test compounds or double distilled water by gavage respectively insingle administration. 30 minutes later, animals were administered with2.5 g/kg glucose by gavage. Before administration and afteradministration of glucose at 30, 60 and 120 minutes, blood was taken tomeasure serum glucose levels.

2. Serum Glucose Determination:

Serum glucose is measured by glucose kits. 250 μl working enzymesolution was taken, and then 5 μl serum was added to the solution. Ablank tube (5 μl double distilled water was added) and a standard tube(5 μl glucose standard solution was added) were establishedsimultaneously and shaken in 37° C. water bath for 20 minutes. The blanktube was used to set the instrument to zero, the colorimetric assay wasperformed at OD 505 nm.Serum glucose concentration (BG, mmol/l)=OD _(sample tube) /OD_(standard tube)×5.55Data Processing and Statistical Analysis:

1. Mean±SD and Student-t test were used in statistical analysis.

2. The percentage of declines in blood glucose after 30 minutes of sugaradministration as well as the area under the curve (AUC) werecalculated.

Results:

Test 1:

Male ICR mice were fasted for 6 hours, and then treated with doubledistilled water, different doses of the test compounds of Example 1 andExample 2 by gavage. 30 minutes after administration, the oral glucosetolerance test was conducted. The results showed that the blood glucoselevel in the control group increased significantly after 2.5 g/kgglucose had been administered by gavage, and reached the peak at 30minutes. At low, middle and high doses of the compound of Example 1, theblood glucose level was significantly lower than the control group at 30minutes, and the percentage of the blood glucose had decreased by19.16%, 22.85% and 31.85%, respectively. At each dose of the compound ofExample 2, the blood glucose level was significantly lower than thecontrol group at 30 minutes after the administration of glucose(P<0.01). Compared with the control group, the percentage of bloodglucose had decreased by 25.54%, 25.92% and 26.93%.

Test 2:

Male ICR mice were fasted for 6 hours, and then treated with doubledistilled water, different doses of the test compounds of Example 1 andExample 2 by gavage. 30 minutes after administration, the oral glucosetolerance test was conducted. The results showed that the blood glucoselevel in the control group increased significantly after 2.5 g/kgglucose had administered by gavage, and reached the peak at 30 minutes.At each dose of SHR1039 (i.e, the compound of Example 1), the bloodglucose level was significantly lower than the control group at 30minutes after the administration of glucose (P<0.01), and the percentageof blood glucose thereof had decreased by 26.10%, 30.24% and 32.05%respectively. At low, middle and high doses of SHR1040 (i.e., thecompound of Example 2), the blood glucose level was significantly lowerthan the control group at 30 minutes (P<0.01), and the percentage ofblood glucose had decreased by 24.51%, 26.96% and 27.75%.

Conclusion:

Two experimental results of this report show that the test compounds ofExample 1 and Example 2 have significant hypoglycemic effect on normalICR mice in the oral glucose tolerance tests. Moreover, the testcompound of Example 1 shows a better dose-effect relationship.

Effects of DPP-IV Inhibitors on Oral Glucose Tolerance in KKAy Mice

Objective:

To observe the DPP-IV inhibitory effects of the compounds of Example 1and Example 2 on oral glucose tolerance tests in type II diabetes KKAymice, a preliminary evaluation of their hypoglycemic effect in vivo wasperformed.

Test Animals:

Species, Strains: KKAy mice

Source: Shanghai Laboratory Animal Center, Chinese Academy of Sciences.Certification No.: SYXK (Shanghai) 2004-2005

Weight: 40˜55 g

Sex: female: 52; male: 33

Raising Conditions: SPF grade animal room raising, temperature: 22-24°C.;

humidity: 45-80%; illumination: 150-300 Lx, day and night cycle with aninterval of 12 hours.

Drugs:

Name: the compounds of Example 1 and Example 2

Preparation Method: Compounds were Weighed Accurately, then Dissolved indouble distilled water, and fully mixed to prepare a 3 mg/mL suspension,then diluted to 1, 0.3, 0.1 mg/mL transparent solutions respectively.

Dose: 1, 3, 10, 30 mg/kg by gavage. The volume is 10 mL/kg.

Methods:

The Effects of the Compounds on Blood Glucose in KKAy Mice

Normal KKAy mice had been fasted for 6 hours, and then were randomlygrouped according to the weights and fasting blood glucose, 5 mice ineach group. The groups included a blank control group as well asdifferent doses of the treatment groups as follows:

Test 1: male 0704

Blank control: double-distilled water by gavage

HR1039: the compound of Example 1, 10 mg/kg by gavage

the compound of Example 1, 30 mg/kg by gavage

Test 2: female 0816

Blank control: double-distilled water by gavage

SHR1039: the compound of Example 1, 3 mg/kg by gavage

the compound of Example 1, 10 mg/kg by gavage

Test 3: male 0712

Black control: double-distilled water by gavage

SHR1040: the compound of Example 2, 3 mg/kg by gavage

the compound of Example 2, 10 mg/kg by gavage

Test 4: female 0907

Black control: double-distilled water by gavage

SHR1040: the compound of Example 2, 3 mg/kg by gavage

the compound of Example 2, 10 mg/kg by gavage

Animals in each group had been fasted for 6 hours, and then pretreatedwith compounds or double distilled water by gavage respectively insingle administration. 30 minutes later, animals were administered 2.5g/kg (female KKAy mice) or 1.5 g/kg (male KKAy mice) glucose by gavage.After administration of glucose at 0, 30, 60 and 120 minutes, serumglucose levels were measured by a glucometer.

Data Processing and Statistical Analysis:

3. Mean±SD and Student-t test, or Anova was used in statisticalanalysis.

4. The percentage of declines in blood glucose after 30 minutes of sugaradministrations and the area under the curve (AUC) were calculated.

Results:

1. Compound of Example 1: Test 1, 2

Male KKAy mice were fasted for 6 hours, and then treated with doubledistilled water and different doses of the test compound of Example 1 bygavage. 30 minutes after administration, the oral glucose tolerance testwas conducted. The results showed that the blood glucose level in thecontrol group increased significantly after 1.5 g/kg glucose had beenadministered by gavage, and reached the peak at 30 minutes. In the dosesof 10 mg/kg and 30 mg/kg of the compound of Example 1 groups, bloodglucose levels were both lower than the control group 30 minutes afterthe administration of glucose. Compared with the control group, thepercentage of blood glucose had decreased by 16.22% and 17.15%respectively.

Female KKAy mice were fasted for 6 hours, and then treated with doubledistilled water and different doses of the test compound of Example 1 bygavage. 30 minutes after administration, the oral glucose tolerance testwas conducted. The results showed that the blood glucose level in thecontrol group increased significantly after 2.5 g/kg glucose hadadministered by gavage, and reached the peak at 30 minutes. In the dosesof 3 mg/kg and 10 mg/kg of the compound of Example 1 groups, bloodglucose levels were both significantly lower than the control group 30minutes after the administration of glucose. The percentage of bloodglucose had decreased by 40.63% and 24.68% respectively.

2. Compound of Example 2: Test 3, 4

Male KKAy mice were fasted for 6 hours, and then treated with doubledistilled water and different doses of the test compound of Example 2 bygavage. 30 minutes after administration, the oral glucose tolerance testwas conducted. The results showed that the blood glucose level in thecontrol group increased significantly after 1.5 g/kg glucose had beenadministered by gavage, and reached the peak at 30 minutes. In the dosesof 10 mg/kg and 30 mg/kg of the compound of Example 2 groups, bloodglucose levels were both lower than the control group 30 minutes afterthe administration of glucose. Compared with the control group, thepercentage of blood glucose had decreased by 13.79% and 12.23%respectively.

Female KKAy mice were fasted for 6 hours, and then treated with doubledistilled water and different doses of the test of compound Example 2 bygavage. 30 minutes after administration, the oral glucose tolerance testwas conducted. The results showed that the blood glucose level in thecontrol group increased significantly after 2.5 g/kg glucose had beenadministrated by gavage, and reached the peak at 30 minutes. In the doseof 10 mg/kg of the compound of Example 2 group, blood glucoses werelower than control group at 30 minutes after the administration ofglucose (P=0.075, ANOVA). The percentage of blood glucose had decreasedby 21.55%. However, since there is a great individual difference in themice, the results had no significant difference.

Conclusion:

The test compounds of Example 1 and Example 2 both have somehypoglycemic effects on oral glucose tolerance test in type II diabetesKKAy mice.

1. A compound of formula (I) or a pharmaceutically acceptable saltthereof:

wherein: R is the following formula:

wherein: R⁵ is selected from the group consisting of hydrogen, alkyl,cycloalkyl, aryl, heteroaryl and heterocyclic alkyl, wherein the alkyl,cycloalkyl, aryl, heteroaryl or heterocyclic alkyl is independentlyunsubstituted or substituted with one or more groups independentlyselected from the group consisting of alkyl, cycloalkyl, aryl,heteroaryl, alkoxyl, cycloalkoxyl, aryloxyl, heteroaryloxyl, halogen,hydroxyl, amino, alkylamino, cyano, hydroxyalkyl, heterocyclic alkyl,heterocyclic alkoxyl, carboxylic acid and carboxylic ester; R⁶ and R⁷are each independently selected from the group consisting of hydrogen,alkyl, aryl, heteroaryl, haloalkyl, haloalkoxyl, hydroxy, amino, cyano,alkynyl, alkoxyl, aryloxyl, hydroxyalkyl, heterocyclic alkyl, carboxylicacid, carboxylic ester and halogen; and W is C, S or O, wherein C isunsubstituted or substituted with R⁶ or R⁷ R¹ is selected from the groupconsisting of —C(O)NR³R⁴, —C(O)R³ and —C(O)OR³; R² is selected from thegroup consisting of hydrogen, unsubstituted or substituted cycloalkyl,and alkyl, wherein the alkyl is unsubstituted or substituted with one ormore groups independently selected from the group consisting ofcycloalkyl and aryl; R³ and R⁴ are either each independently selectedfrom the group consisting of hydrogen, alkyl, cycloalkyl, aryl,heteroaryl and heterocyclic alkyl, wherein the alkyl, cycloalkyl, aryl,heteroaryl or heterocyclic alkyl is independently unsubstituted orsubstituted with one or more groups independently selected from thegroup consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxyl,cycloalkoxyl, aryloxyl, heteroaryloxyl, halogen, hydroxyl, amino, cyano,hydroxyalkyl, heterocyclic alkyl, heterocyclic alkoxyl, trifluoromethyl,carboxylic acid and carboxylic ester; or R³ and R⁴, together with the Natom to which they are attached, form a 3- to 8-membered heterocyclicring, wherein the 3- to 8-membered heterocyclic ring further containszero or more heteroatoms selected from N, O and S, and the 3- to8-membered ring is unsubstituted or substituted with one or more groupsindependently selected from the group consisting of alkyl, aryl,heteroaryl, haloalkyl, haloalkoxyl, hydroxyl, amino, cyano, alkoxyl,aryloxyl, hydroxyalkyl, heterocyclic alkyl, carboxylic acid, carboxylicester, and halogen; and n is an integer from 0 to
 4. 2. The compound ofclaim 1, or a pharmaceutically acceptable salt thereof, which is thecompound of formula (IA) or a pharmaceutically acceptable salt thereof:

wherein: R is the following formula:

wherein: R⁵ is selected from the group consisting of hydrogen, alkyl,cycloalkyl, aryl, heteroaryl and heterocyclic alkyl, wherein the alkyl,cycloalkyl, aryl, heteroaryl or heterocyclic alkyl is independentlyunsubstituted or substituted with one or more groups independentlyselected from the group consisting of alkyl, cycloalkyl, aryl,heteroaryl, alkoxyl, cycloalkoxyl, aryloxyl, heteroaryloxy, halogen,hydroxyl, amino, alkylamino, cyano, hydroxyalkyl, heterocyclic alkyl,heterocyclic alkoxyl, carboxylic acid and carboxylic ester; R⁶ and R⁷are each independently selected from the group consisting of hydrogen,alkyl, aryl, heteroaryl, haloalkyl, haloalkoxyl, hydroxyl, amino, cyano,alkynyl, alkoxyl, aryloxyl, hydroxyalkyl, heterocyclic alkyl, carboxylicacid, carboxylic ester and halogen; and W is C, S or O, wherein C isunsubstituted or substituted with R⁶ or R⁷ R¹ is selected from the groupconsisting of —C(O)NR³R⁴, —C(O)R³ and —C(O)OR³; R² is selected from thegroup consisting of hydrogen, unsubstituted or substituted cycloalkyl,and alkyl, wherein the alkyl is unsubstituted or substituted with one ormore groups independently selected from the group consisting ofcycloalkyl and aryl; R³ and R⁴ are either each independently selectedfrom the group consisting of hydrogen, alkyl, cycloalkyl, aryl,heteroaryl and heterocyclic alkyl, wherein the alkyl, cycloalkyl, aryl,heteroaryl or heterocyclic alkyl is independently unsubstituted orsubstituted with one or more groups independently selected from thegroup consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxyl,cycloalkoxyl, aryloxyl, heteroaryloxyl, halogen, hydroxyl, amino, cyano,hydroxyalkyl, heterocyclic alkyl, heterocyclic alkoxyl, trifluoromethyl,carboxylic acid and carboxylic ester; and or R³ and R⁴, together withthe N atom to which they are attached, may form a 3- to 8-memberedheterocyclic ring, wherein the 3- to 8-membered heterocyclic ringcontains zero or more heteroatoms selected from N, O and S, and the 3-to 8-membered ring is unsubstituted or substituted with one or moregroups independently selected from the group consisting of alkyl, aryl,heteroaryl, haloalkyl, haloalkoxyl, hydroxyl, amino, cyano, alkoxyl,aryloxyl, hydroxyalkyl, heterocyclic alkyl, carboxylic acid, carboxylicester, and halogen.
 3. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, which is the compound of formula (IB) or apharmaceutically acceptable salt thereof:

wherein: R is the following formula:

R¹ is selected from the group consisting of —C(O)NR³R⁴, —C(O)R³ and—C(O)OR³; R² is selected from the group consisting of hydrogen,unsubstituted or substituted cycloalkyl, and alkyl, wherein the alkyl isunsubstituted or substituted with one or more groups independentlyselected from the group consisting of cycloalkyl and aryl; R³ and R⁴ areeither each independently selected from the group consisting ofhydrogen, alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic alkyl,wherein the alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic alkyl isindependently unsubstituted or substituted with one or more groupsindependently selected from the group consisting of alkyl, cycloalkyl,aryl, heteroaryl, alkoxyl, cycloalkoxyl, aryloxyl, heteroaryloxyl,halogen, hydroxyl, amino, cyano, hydroxyalkyl, heterocyclic alkyl,heterocyclic alkoxyl, trifluoromethyl, carboxylic acid and carboxylicester; or R³ and R⁴, together with the N atom to which they areattached, may form a 3- to 8-membered heterocyclic ring, wherein the 3-to 8-membered heterocyclic ring contains zero or more heteroatomsselected from N, O and S, and the 3- to 8-membered ring is unsubstitutedor substituted with one or more groups independently selected from thegroup consisting of alkyl, aryl, heteroaryl, haloalkyl, haloalkoxyl,hydroxyl, amino, cyano, alkoxyl, aryloxyl, hydroxyalkyl, heterocyclicalkyl, carboxylic acid, carboxylic ester, and halogen; R⁵ is selectedfrom the group consisting of hydrogen, alkyl, cycloalkyl, aryl,heteroaryl and heterocyclic alkyl, wherein the alkyl, cycloalkyl, aryl,heteroaryl or heterocyclic alkyl is independently unsubstituted orsubstituted with one or more groups independently selected from thegroup consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxyl,cycloalkoxyl, aryloxyl, heteroaryloxyl, halogen, hydroxyl, amino,alkylamino, cyano, hydroxyalkyl, heterocyclic alkyl, heterocyclicalkoxyl, carboxylic acid and carboxylic ester; R⁶ and R⁷ are eachindependently selected from the group consisting of hydrogen, alkyl,aryl, heteroaryl, haloalkyl, haloalkoxyl, hydroxyl, amino, cyano,alkynyl, alkoxyl, aryloxyl, hydroxyalkyl, heterocyclic alkyl, carboxylicacid, carboxylic ester and halogen; and W is C, S or O, wherein C isunsubstituted or substituted with R⁶ or R⁷.
 4. The compound of claim 1,or a pharmaceutically acceptable salt thereof, which is the compound offormula (IC) or a pharmaceutically acceptable salt thereof:

wherein: R is the following formula:

R¹ is selected from the group consisting of —C(O)NR³R⁴, —C(O)R³ and—C(O)OR³; R² is selected from the group consisting of hydrogen,unsubstituted or substituted cycloalkyl, and alkyl, wherein the alkyl isindependently unsubstituted or substituted with one or more groupsindependently selected from the group consisting of cycloalkyl and aryl;R³ and R⁴ are each independently selected from the group consisting ofhydrogen, alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic alkyl,wherein the alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic alkyl isindependently unsubstituted or substituted with one or more groupsindependently selected from the group consisting of alkyl, cycloalkyl,aryl, heteroaryl, alkoxyl, cycloalkoxyl, aryloxyl, heteroaryloxyl,halogen, hydroxyl, amino, cyano, hydroxyalkyl, heterocyclic alkyl,heterocyclic alkoxyl, trifluoromethyl, carboxylic acid and carboxylicester; or R³ and R⁴, together with the N atom to which they areattached, may form a 3- to 8-membered heterocyclic ring, wherein the 3-to 8-membered heterocyclic ring contains zero or more heteroatomsselected from N, O and S, and the 3- to 8-membered ring is unsubstitutedor substituted with one or more groups independently selected from thegroup consisting of alkyl, aryl, heteroaryl, haloalkyl, haloalkoxyl,hydroxyl, amino, cyano, alkoxyl, aryloxyl, hydroxyalkyl, heterocyclicalkyl, carboxylic acid, carboxylic ester, and halogen; R⁵ is selectedfrom the group consisting of hydrogen, alkyl, cycloalkyl, aryl,heteroaryl and heterocyclic alkyl, wherein the alkyl, cycloalkyl, aryl,heteroaryl or heterocyclic alkyl is independently unsubstituted orsubstituted with one or more groups independently selected from thegroup consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxyl,cycloalkoxyl, aryloxyl, heteroaryloxyl, halogen, hydroxyl, amino,alkylamino, cyano, hydroxyalkyl, heterocyclic alkyl, heterocyclicalkoxyl, carboxylic acid and carboxylic ester; R⁶ and R⁷ are eachindependently selected from the group consisting of hydrogen, alkyl,aryl, heteroaryl, haloalkyl, haloalkoxyl, hydroxyl, amino, cyano,alkynyl, alkoxyl, aryloxyl, hydroxyalkyl, heterocyclic alkyl, carboxylicacid, carboxylic ester and halogen; and W is C, S or O, wherein C isunsubstituted or substituted with R⁶ or R⁷.
 5. The compound of claim 1,or a pharmaceutically acceptable salt thereof, wherein the salt isselected from the group consisting of a hydrochloride salt, ap-toluenesulfonate salt, a tartrate salt, a malate salt, a lactate salt,a methanesulfonate salt, a sulfate salt, a phosphate salt, a citratesalt, an acetate salt, and a trifluoroacetate salt.
 6. The compound ofclaim 5, or a pharmaceutically acceptable salt thereof, wherein the saltis selected from the group consisting of a hydrochloride salt, ap-toluenesulfonate salt, a tartrate salt, and a trifluoroacetate salt.7. The compound of claim 1, or a pharmaceutically acceptable saltthereof, which is selected from the group consisting of:


8. A pharmaceutical composition comprising a therapeutically effectiveamount of the compound of claim
 1. 9. A process for preparing thecompound of formula (IB) of claim 3, comprising the step of:

converting the compound of formula (ID) the compound of formula (IB).10. A process of claim 9, wherein the converting is carried out inmethanol or ethanol.
 11. A process for preparing the compound of formula(IC) of claim 4, comprising the steps of:

converting the compound of formula (ID) to an azabicyclo alkenylcompound;

converting the azabicyclo alkenyl compound an azabicyclo cyano compound;

converting the azabicyclo cyano compound to an azabicyclo aminecompound; and

reacting the azabicyclo amine compound with a halo substituted compoundto obtain the compound of formula (IC).
 12. A process of claim 11,wherein the converting of the azabicyclo alkenyl compound is carried outin the presence of dichloromethane, and the converting of the azabicyclocyano compound is carried out in the presence of ethanol andhydrochloric acid.
 13. A process for preparing the compound of formula(IB) of claim 3, comprising the steps of:

converting the compound of formula (ID) to an azabicyclo cyano compound;

converting the azabicyclo cyano compound to a R² substituted azabicyclocyano compound;

converting the R² substituted azabicyclo cyano compound to a R²substituted azabicyclo carboxyl compound;

converting the R² substituted azabicyclo carboxyl compound a substitutedazabicyclo azido compound;

converting the azabicyclo azido compound to a R² substituted azabicycloamine compound; and

converting the R² substituted azabicyclo amine compound to the compoundof formula (IB).
 14. A process for preparing the compound of formula(IC) of claim 4, comprising the following steps of:

reducing the intermediate of formula (ID) to an azabicyclo hydroxylcompound;

converting the azabicyclo hydroxyl compound to an azabicyclo methylsulfonic acid compound;

converting the azabicyclo methyl sulfonic acid compound to a phthalimidesubstituted azabicyclo compound;

converting the phthalimide substituted azabicyclo compound to anazabicyclo amine compound; and

converting the azabicyclo amine compound to the compound of formula(IC).
 15. A process of claim 14, wherein the reducing is carried out inthe presence of lithium tri-tert-butoxyaluminum hydride.
 16. Apreparation process of claim 11 or 13, further comprising the step ofreacting the compound of formula (IB) or formula (IC) with an acid toobtain an acid addition salt.
 17. A method of inhibiting a dipeptidylpeptidase comprising administering the compound or pharmaceuticallyacceptable salt thereof of claim 1 to a patient in need of thetreatment.
 18. A method of inhibiting a dipeptidyl peptidase comprisingadministering the composition of claim 8 to a patient in need of thetreatment.
 19. The compound of claim 1, or a pharmaceutically acceptablesalt thereof, wherein R³ and R⁴ are independently selected from thegroup consisting of hydrogen, alkyl, and hydroxylalkyl.
 20. The compoundof claim 1, or a pharmaceutically acceptable salt thereof, wherein R¹ is—C(O)NR³R⁴.
 21. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein R¹ is selected from the groupconsisting of —C(O)N(CH₃)₂, —C(O)NCH₂(CH₃)₂, —C(O)CH₃, —C(O)OCH₃, and—C(O)CH₂OH.
 22. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein R² is selected from the groupconsisting of hydrogen, and unsubstituted or substituted alkyl.
 23. Thecompound of claim 1, or a pharmaceutically acceptable salt thereof,wherein R² is selected from the group consisting of hydrogen, methyl,ethyl, cyclohexylmethyl, cyclopentyl, and benzyl.
 24. The compound ofclaim 23, or a pharmaceutically acceptable salt thereof, wherein R² ismethyl or ethyl.
 25. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein R⁵ is hydrogen.
 26. The compound ofclaim 1, or a pharmaceutically acceptable salt thereof, wherein at leastone of R⁶ and R⁷ is hydrogen.
 27. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein at least one of R⁶ andR⁷ is cyano.
 28. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein W is C.
 29. The compound of claim 28,or a pharmaceutically acceptable salt thereof, wherein W ishalogen-substituted C.
 30. The compound of claim 1, or apharmaceutically acceptable salt thereof, which is selected from thegroup consisting of:


31. The preparation process of claim 13, further comprising the step ofreacting the compound of formula (IB) with an acid to obtain an acidaddition salt.
 32. A method of inhibiting dipeptidyl peptidase-IV in amammal in need thereof comprising the step of administering atherapeutically effective amount of the compound of claim 1, or apharmaceutically acceptable salt thereof.
 33. A method of treating typeII diabetes comprising administering to a mammal in need thereof atherapeutically effective amount of the compound of claim 1, or apharmaceutically acceptable salt thereof.